Patent Application
20220313829
The present invention is in the field of medicinal chemistry and particularly relates to a bifunctional compound, preparation method and use thereof. The bifunctional compound can be used to prevent or treat cancers, especially cancers harboring abnormal expression of EGFR, Her2, Her3 or Her4 proteins etc.
Lung cancer is the leading cause of cancer death and the most commonly diagnosed cancer not only in China but in the worldwide. According to the latest epidemiological data of the Chinese oncology in 2019, there are 572.6 thousand new cases and 458.7 thousand deaths of lung cancer each year in China. Currently, the 5-year survival rate of lung cancer is only 17%, which has changed little since 1970s. The most important reason is that the conventional radiotherapy and chemotherapy with strong toxic and side effects, which not only kill cancer cells, but also normal cells in patients, could not prevent the progress of lung cancer. It's urgent to explore a new way for improving the survival rate and quality of life of patients with lung cancer.
Targeted therapy can reduce the toxic and side effects compared to chemotherapy or radiotherapy in patients harboring special oncogenes. The use of tyrosine kinase inhibitors (TKIs), for instance, can increase the therapy effect from 40% of the chemotherapy up to 70%, and the PFS (progress free survival) will raise to about 10 months from 5 months in patients with aberrant epidermal growth factor receptor (EGFR). EGFR is one of the family members of epidermal growth factor receptor tyrosine kinase, and the morbidity of lung cancers with abnormal EGFR expression is higher amongst eastern populations which accounts for 50% of lung adenocarcinoma, than that in western populations with the morbidity of 15%, which means that the targeted drugs for EGFR will benefit Asian patients well. However, almost all of the patients will develop resistance to the targeted drugs at about 1 year after treatment, causing tumor progress again. Researches on the drug resistant mechanisms against the 1st and 2nd EGFR TKIs showed that about 60% of these lung cancer patients acquired the secondary EGFR mutation T790M. These findings promote the development of the next EGFR targeted drugs and are expected to improve the life quality of patients. The first generation EGFR TKIs such as Erlotinib (trade name “Tarceva”) and Gefitinib (trade name “Iressa”), which can combine with EGFR tyrosine kinase domain on the ATP binding pocket reversibly, and the second generation irreversible EGFR TKI Afatinib (approved by FDA in July 2013), have been approved by FDA for clinical use at present. The second generation EGFR TKI Dacomitinib, approved by FDA in 2018, was used for the first line treatment on the locally advanced or metastatic non-small cell lung cancer (NSCLC) patients harboring EGFR activation mutations. The third generation EGFR TKI Osimertinib (AZD9291), approved by FDA at the end of 2015, can specifically kill cancer cells with EGFR activating mutations (EGFR exon 19del or EGFR exon 20 L858R) and T790M, the drug resistance mutation, and can extend the PFS of the lung cancer patients with T790M mutation about one year. However, research found that the acquired resistance against AZD9291 was developed inevitably due to some complicated reasons including the incidence of the tertiary mutation C797S. The recurrence of tumors and their resistance to targeted drugs indicated that the small molecular inhibitors, without a long-term efficacy, couldn't improve the quality of survival for patients or meet the need of social development. Patients of lung cancer with EGFR mutations account for a high proportion of Chinese lung cancer patients, and the rate are still increasing year by year. Thus, it's necessary to explore new therapies and drugs for lung cancers to overcome drug resistance problems on small molecular targeted compounds.
We have developed a brand-new technology for targeted drugs called PROTAD:PROteolysis TArgeting Drug, which aimed to change the fate of those disease proteins by using the ubiquitination/proteasome system, the intracellular protein degradation machine. PROTAD is composed of two ligands, one of which can target the disease protein and the other can bind with ubiquitin-protein ligase (E3), connected by a linker. The bi-functional small molecular can compel the proteins of interest ubiquitinated, and then transfer them to the degradation machine. Compared with the traditional small molecule drug design, the biggest difference of PROTAD is that it mobilizes the whole cell as the drug effector unit. The PROTAD only need a transient combination with targets to tag the proteins as “to be cleaned”, and then the compounds can be recycled, which means can a low PROTAD dosage not only meet the need of therapy, but reduce the off-target risk. Due to the above advantages, PROTAD, potential to eliminate the tumor progress caused by oncogenes and the acquired drug resistance, is expected to conquer the difficulties in tumor targeted therapy.
It is an objective of the present invention to provide a bifunctional compound, preparation method and use thereof to overcome the above mentioned disadvantages and solve the problems in the prior art.
In order to achieve the above-mentioned objectives and other related objectives, in one aspect of the present invention, there is provided a bifunctional compound of Formula I:
or a pharmaceutically acceptable salt, an isomer, a prodrug, a polymorph, or a solvate thereof, wherein
EGFR Binders can bind to EGFR protein;
ULM represents:
or, ULM represents:
or, ULM represents:
LIN represents a linking group covalently bonded to the EGFR Binders and ULM, respectively.
In another aspect of the present invention, there is provided the use of the bifunctional compound in the manufacture of a medicament.
In another aspect of the present invention, there is provided a pharmaceutical composition, comprising the bifunctional compound or a pharmaceutically acceptable salt, an isomer, a prodrug, a polymorph, or a solvate thereof, and at least one pharmaceutically acceptable carrier, an additive, an adjuvant, or an excipient.
As a result of extensive studies, the present inventors synthesized and developed a new class of bifunctional compounds based on different EGFR-related drugs, e.g., EGFR inhibitors, e.g., Dacomitinib, Poziotinib, Gefitinib, Afatinib, Sapitinib, Canertinib, Osimertinib, and EAI045, etc. The bifunctional compounds of the present invention show different degrees of regulatory effects on EGFR protein, which can not only promote the degradation of EGFR protein, but also inhibit the activity of EGFR kinase and the proliferation of EGFR mutation-positive cells, and thus can be used as a therapeutic agent for tumor patients. In view of the above, the present invention has been completed.
Designing degraders that target specific proteins is a new mode of drug development. In the present invention, the present inventors designed special bispecific protein modulators by using the Proteolysis Targeting Drug (PROTAD) technology platform, which can tag the target proteins as “to be degraded”, and degrade them by activating the protein degradation pathway inside the cell. Compared with traditional small molecule drug design, the proteins-targeted small molecule modulators of the present invention can induce the degradation of the target proteins, which is essentially different from the traditional small molecule inhibitor in mechanism.
In one aspect of the present invention, there is provided a bifunctional compound of Formula I:
or a pharmaceutically acceptable salt, an isomer, a prodrug, a polymorph, or a solvate thereof, wherein
EGFR Binders can bind to EGFR protein;
ULM represents:
or, ULM represents:
or, ULM represents:
Unless otherwise specified, the isotope-labeled forms of the compounds of the present invention are also encompassed within the scope of the present invention. For example, in the compounds of the present invention with the structure/formula given above, at least one hydrogen atom is replaced by deuterium or tritium, or at least one carbon atom is replaced by 13C- or 14C-enriched carbon, or at least one nitrogen atom is replaced by 15N-enriched nitrogen.
In the present invention, the term “salt” should be understood as any form of active compounds used in the present invention, wherein the compounds may be in ionic form or charged or coupled to a counter-ion (cation or anion) or in solution. The term “salt” can also include quaternary ammonium salts and complexes of the active compounds with other molecules and ions, especially complexes through ionic interactions. The term “salt” especially includes physiologically acceptable salts, and can be understood to be equivalent to “pharmacologically acceptable salts”.
In the present invention, the term “pharmaceutically acceptable salt” generally refers to any salt that is physiologically tolerable (generally speaking, this means that it is non-toxic, especially as a result of counter-ion is non-toxic) when used in a suitable manner for treatment (especially when applied or used in humans and/or mammals). These physiologically acceptable salts may be formed with cations or bases, and in the context of the present invention, especially when administered in humans and/or mammals, they should be understood to be a salt formed by at least one compound provided in accordance with the invention (usually a (deprotonated) acid), such as an anion, and at least one physiologically tolerable cation (preferably an inorganic cation). In the context of the present invention, it may specifically include salts formed with alkali metals and alkaline earth metals, and salts formed with ammonium cations (NH4+), specifically including but not limited to (mono) or (di) sodium, (mono) or (di) potassium, magnesium or calcium salts. These physiologically acceptable salts can also be formed with anions or acids, and in the context of the present invention, especially when administered in humans and/or mammals, they should be understood as a salt formed by at least one compound provided in accordance with the present invention (usually protonated (for example on nitrogen)), such as a cation and at least one physiologically tolerable anion. In the context of the present invention, it may specifically include a salt formed by a physiologically tolerable acid, that is, a salt formed by a specific active compound and a physiologically tolerable organic or inorganic acid, and specifically may include but not limited to salts formed with hydrochloric acid, hydrobromic acid, sulfuric acid, methanesulfonic acid, formic acid, acetic acid, oxalic acid, succinic acid, malic acid, tartaric acid, mandelic acid, fumaric acid, lactic acid, or citric acid.
The compound of formula I of the present invention may include an enantiomer depending on the presence of a chiral center or an isomer depending on the presence of a double bond (for example, Z, E). Single isomer, enantiomers, diastereomers, or cis-trans isomers, and mixtures thereof are also encompassed within the scope of the present invention.
In the present invention, the term “prodrug” is used in its broadest sense and includes those derivatives that can be converted into the compounds of the present invention in vivo. Methods for preparing the prodrugs of a designated active compound should be known to those skilled in the art. For example, one can refer to related content disclosed in Krogsgaard-Larsen et al., “Textbook of Drug design and Discovery”, published by Taylor & Francis (April 2002).
In the present invention, the term “solvate” generally refers to any form of the active compound according to the present invention bonded to another molecule (usually a polar solvent) through a non-covalent bond, and the obtained substance may specifically include but not limited to hydrates and alcoholates, such as methanolates.
The bifunctional compounds of the present invention can comprise an EGFR Binders moiety, which is usually covalently bonded to LIN and can usually bind to EGFR protein. In the present invention, the EGFR Binders can be any molecule that can bind to EGFR protein, more specifically EGFR TKIs. The EGFR TKIs (epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors) can act on intracellular protein tyrosine kinase domain of EGFR; the epidermal growth factor receptor tyrosine kinase inhibitor can usually bind to the tyrosine kinase functional domain competitively with ATP, and can reversibly or irreversibly inhibit tyrosine kinase phosphorylation. The EGFR TKIs part is usually used as the protein target binding moiety (PTM, protein target moiety), which can be linked to the ULM moiety (E3 ubiquitin ligase binding moiety) via LIN, thereby leading to the ubiquitination of the target proteins, and the activation of the intracellular proteasome system for targeted degradation of the target proteins. The ubiquitination degradation pathway can degrade most of the ubiquitinated proteins in the cells, e.g., 80% to 90% or higher of the ubiquitinated proteins in the cells. If this system can be activated to specifically clean up the carcinogen proteins, which restores the cellularprotein homeostasis, it is likely to alleviate or treat cancers. The PROTAD technology takes advantage of this, and uses the specially designed dual-specific degraders to tag the target proteins as “to be ubiquitined” to achieve targeted degradation.
In the bifunctional compounds of the present invention, the EGFR TKIs can specifically represents a group as shown by the following formula:
wherein R1, R2, R3, and R4 are each independently selected from H, halogen, Cl, F, C1-10 alkyl, C1-10 haloalkyl, C1-10 alkynyl, C1-10 alkoxy, arylmethoxy, and heteroarylmethoxy, wherein the aryl of the arylmethoxy and the heteroaryl of the heteroarylmethoxy are unsubstituted or are substituted by 1-2 substituent(s) selected from C1-10 alkyl, halogen, and C1-10 haloalkyl;
one of R5 and R6 is covalently bonded to LIN, and forms —NR″—, where R″ is selected from H, linear or branched C1-C10 alkyl, or C3-C9 cycloalkyl, or forms a group as shown by the following formulas:
one of R5 and R6 is selected from H, N, halogen, C1-10alkyl, C1-10 haloalkyl, C1-10 alkoxy, amino, acylamino, alkylamino, di-C1-10alkylamino, cyano, aryl, heteroaryl, furyl, pyrrolyl, imidazolyl, oxazolyl, isoxazolyl, triazolyl, C3-9 cycloalkyl, C3-9 cycloalkyloxy, heterocyclyl, heterocyclyloxy, and —NHC(O)R14, wherein the aryl and the heteroaryl are unsubstituted or are substituted by 1-2 substituent(s) selected from C1-10 alkyl, halogen, C1-10haloalkyl, cyano, R7SO2(CH2)sNHCH2—, —OR8, and —NHC(O)R9, wherein R7 is selected from C1-10 alkyl and s is 0, 1, 2, or 3; and R8 is selected from C1-10 alkyl which is optionally mono- or multi-substituted by the groups independently selected from hydroxyl, C1-10 alkoxy, amino, C1-10 alkylamino, and di-C1-10 alkylamino; and R9 is selected from the following groups:
In the present invention, the term “halogen” or “halo” or “halogenated” generally refers to fluorine, chlorine, bromine or iodine.
In the present invention, the term “alkyl” generally refers to saturated aliphatic groups, which can be linear or branched. For example, C1-10 alkyl generally refers to an alkyl group including 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms, and may specifically include, but is not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, etc. Again for example, C1-30 alkylene generally refers to an alkylene group including 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 carbon atoms, and may specifically include, but is not limited to, methylene, ethylene, propylene, butylene, pentylene, hexylene, heptylene, octylene, nonylene, and decylene, etc.
In the present invention, the term “haloalkyl” generally refers to halogenated saturated aliphatic groups, which can be linear or branched, and are optionally independently mono- or multi-substituted by the group selected from fluorine, chlorine, bromine, or iodine. For example, C1-10 haloalkyl generally refers to a haloalkyl group including 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms, and may specifically include, but is not limited to, halomethyl, haloethyl, halopropyl, halobutyl, halopentyl, halohexyl, haloheptyl, halooctyl, halononyl, halodecyl, etc.
In the present invention, the term “C3-9 cycloalkyl” generally refers to a saturated or unsaturated (but not aromatic) cyclic hydrocarbon having from 3 to 9 carbon atoms. The cycloalkyl group may specifically include, but is not limited to, cyclopropyl, 2-methylcyclopropyl, cyclopropylmethyl, cyclobutyl, cyclopentyl, cyclopentylmethyl, cyclohexyl, cycloheptyl, cyclooctyl, adamantanyl, noradamantanyl, etc.
In the present invention, the term “heterocyclyl” generally refers to a saturated or unsaturated (but not aromatic) cyclic hydrocarbon, containing at least one heteroatoms selected from N, O or S. The heterocyclyl group may specifically include, but are not limited to, pyrrolinyl, pyrrolidinyl, pyrazolinyl, aziridinyl, azetidinyl, tetrahydropyrrolyl, oxiranyl, oxetanyl, dioxetanyl, tetrahydropyranyl, tetrahydrofuranyl, dioxanyl, dioxolanyl, oxazolidinyl, piperidinyl, piperazinyl, morpholinyl, azepanyl or diazepanyl, etc. Preferably, the heterocyclyl group in the present invention is usually a 5- or 6-membered ring system.
In the present invention, the term “aryl” generally refers to a group having at least one aromatic ring but no heteroatoms. The aryl group may optionally be mono- or multi-substituted with substituents independently selected from alkyl, halogen, haloalkyl, cyano, R7SO2(CH2)sNHCH2—, —OR8, —NHC(O)R9. The aryl group may specifically include, but is not limited to, phenyl, naphthyl, fluoranthenyl, fluorenyl, tetrahydronaphthyl, indanyl, anthracyl, etc. Preferably, the aryl group in the present invention is a 5- or 6-membered ring system which is optionally at least monosubstituted.
In the present invention, the term “heteroaryl” generally refers to a heterocyclic ring system having at least one aromatic ring and optionally containing one or more heteroatoms selected from N and O, and may optionally be mono- or multi-substituted by the substituents independently selected from alkyl, halogen, haloalkyl, cyano, R7SO2(CH2)sNHCH2—, —OR8, and —NHC(O)R9. The heteroaryl group may specifically include, but is not limited to, furanyl, benzofuranyl, pyrrolyl, pyridinyl, pyrimidinyl, pyridazinyl, pyrazinyl, quinolinyl, isoquinolinyl, phthalazinyl, triazolyl, pyrazolyl, isoxazolyl, indolyl, benzotriazolyl, benzodioxolanyl, benzodioxanyl, benzimidazolyl, carbazolyl, quinazolinyl, etc. Preferably, the heteroaryl group in the present invention is a 5- or 6-membered ring system which is optionally at least mono-substituted.
In the present invention, the term “alkenyl” generally refers to an unsaturated aliphatic group including at least one C═C double bond. The alkenyl group may specifically include, but is not limited to, vinyl, propenyl, butenyl, etc.
In some preferred embodiments of the present invention, in the formula V, R1, R2, R3, and R4 are each independently selected from H, halogen, Cl, and F.
In a more preferred embodiment of the present invention, in the formula V, R1 is selected from Cl, R2 is selected from F, R3 and R4 are each independently selected from H; or, R1 is selected from H, R2 is selected from Cl, R3 is selected from Cl, and R4 is selected from F.
In some preferred embodiments of the present invention, in the formula V, one of R5 and R6 may be covalently bonded to LIN, and form —NR″—, where R″ is selected from H, linear or branched C1-C10 alkyl, or C3-C10 cycloalkyl; or form a group as shown by the following formulas:
one of R5 and R6 is selected from C1-C10 alkoxy, heterocyclyloxy, and —NHC(O)R14, wherein R14 is selected from C1-C10 alkyl and alkenyl.
In a more preferred embodiment of the present invention, in the formula V, R5 can be covalently bonded to LIN, and forms —NH—, or forms a group as shown by the following formulas:
In a more preferred embodiment of the present invention, in the formula V, R6 may be selected from methoxy, or a group as shown by the following formula:
In a more preferred embodiment of the present invention, in the formula V, R5 can be selected from —NHC(O)R14, where R14 is selected from vinyl.
In a more preferred embodiment of the present invention, in the formula V, R6 can be covalently bonded to LIN, and represents a group shown by one of the following formulas:
In a further preferred embodiment of the present invention, the EGFR TKIs represents a group shown by one of the following formulas:
In the bifunctional compounds of the present invention, the EGFR TKIs can also specifically represent the group shown by the following formula:
In some preferred embodiments of the present invention, in the formula VI, R16, R17, R18, R19, and R20 are each independently selected from H, OH, and F;
In a more preferred embodiment of the present invention, in the formula VI, R16 is selected from H and OH; R17 is selected from H; R18 is selected from H; R19 is selected from H and F; R20 is selected from H;
In some preferred embodiments of the present invention, R15 is covalently bonded to LIN, and forms a group selected from ones shown by the following formulas:
In a further preferred embodiment of the present invention, the EGFR TKIs represent a group shown by the following formulas:
In the bifunctional compounds of the present invention, the EGFR TKIs can also specifically represent a group shown by the following formula:
In some preferred embodiments of the present invention, in the formula VII, one of R21 and R22, which is covalently bonded to LIN, forms —NR′″—, where R′″ is selected from H, linear or branched C1-C10 alkyl, or C3-C9 cycloalkyl; or forms a group as shown by the following formulas:
In a further preferred embodiment of the present invention, the EGFR TKIs represents a group as shown by one of the following formulas:
The bifunctional compound of the present invention may include a ULM moiety usually covalently bonded to LIN, which is mainly used to bind to E3 ubiquitin ligase as a ligand of E3 ubiquitin ligase.
In some preferred embodiments of the present invention, ULM can represent a group as shown by the following formula:
In a more preferred embodiment of the present invention, in the formula II, A is selected from —CH2— and —(C═O)—; B is selected from C and N; and X, Y, and Z are each independently selected from CH and N;
In a more preferred embodiment of the present invention, in the formula II, R is selected from —S—, —NH—, and ethynylene; or R is absent; D is selected from —(C═O)—; or D is absent.
In a further preferred embodiment of the present invention, ULM can represent a group as shown by the following formulas:
In some preferred embodiments of the present invention, ULM can represent a group as shown by the following formula:
In a more preferred embodiment of the present invention, in the formula III, Z is selected from —(C═O)—.
In a further preferred embodiment of the present invention, ULM can represent a group as shown by the following formula:
In some preferred embodiments of the present invention, ULM can represent a group as shown by the following formula:
The bifunctional compound of the present invention may include a LIN moiety usually covalently bonded to the EGFR TKIs moiety and the ULM moiety, which is mainly used to connect two key parts, namely the target protein and the ligand of E3 ubiquitin ligase. The length, type, and hydrophobicity of the LIN moiety usually have an impact on the stability of the finally formed target protein-PROTAD-E3 ligase ternary complex, which in turn affects its inhibitory and degradation activities. The LIN moiety suitable for connecting PTM (protein target binding portion) and ULM (E3 ubiquitin ligase binding moiety) should be known to those skilled in the art (see, e.g., contents described in Org. Lett. 2019, 21, 3838-3841; and Bioorg. Med. Chem. Lett. 2016; 26:5260-5262). In the present invention, the LIN can specifically represent:
—W-alkylene-;
In a more preferred embodiment of the present invention, the substituents of the linear or branched alkylene are each independently selected from hydroxyl, amino, mercapto, and halogen.
In a more preferred embodiment of the present invention, the alkylene is C1-30 alkylene.
In a more preferred embodiment of the present invention, the LIN represents:
In a further preferred embodiment of the present invention, the LIN represents:
In a further preferred embodiment of the present invention, the LIN represents:
In a further preferred embodiment of the present invention, the LIN represents: —W—(CH2)3CH(OH)CH(OH)(CH2)4—.
In a more preferred embodiment of the present invention, the LIN represents:
In a further preferred embodiment of the present invention, the LIN represents:
In a further preferred embodiment of the present invention, the LIN represents:
In a further preferred embodiment of the present invention, the LIN represents:
In a further preferred embodiment of the present invention, the LIN represents: —W—(CH2)4NHCONH(CH2)4—.
In a further preferred embodiment of the present invention, the LIN represents: —W—(CH2)5S(CH2)5—, —W—(CH2)6S(CH2)5—.
In a further preferred embodiment of the present invention, the LIN represents: —W—(CH2)5SO(CH2)5—, —W—(CH2)6SO(CH2)5—.
In a further preferred embodiment of the present invention, the LIN represents: —W—(CH2)5SO2(CH2)5—, —W—(CH2)6SO2(CH2)5—.
In a further preferred embodiment of the present invention, the LIN represents: —W—(CH2)4CH═CH(CH2)3—.
In a further preferred embodiment of the present invention, the LIN represents: —W—(CH2)2C≡C(CH2)2—, —W—(CH2)5C≡C(CH2)4—.
In a further preferred embodiment of the present invention, the LIN represents:
In a further preferred embodiment of the present invention, the LIN represents:
In a further preferred embodiment of the present invention, the LIN represents: —W-piperazinylene-, —W-spirocycloalkylene, —W-phenylene-, —W—C≡C—C≡C—. Specifically, the LIN can be a group as shown by the following formulas:
In a further preferred embodiment of the present invention, the bifunctional compound is selected from the compounds shown in Table 1 or Table 2:
In a second aspect of the present invention, there is provided the use of the bifunctional compounds provided in the first aspect of the present invention in the preparation of a medicament. As mentioned above, the bifunctional compounds of the present invention include the EGFR TKIs moiety and the ULM moiety, which are covalently linked to LIN, respectively. The EGFR TKIs moiety is usually used as a protein target binding moiety (PTM, protein target moiety), which (epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors) can act on the intracellular protein tyrosine kinase domain of EGFR, and the ULM moiety can compel the target protein ubiquitinated, thereby inducing the degradation of the target protein by activating the intracellular proteasome system. The ubiquitination degradation pathway can degrade most of the ubiquitinated proteins in the cells. For example, it can degrade 80% to 90% or higher of the ubiquitinated proteins in the cells. If this system can be activated to specifically clean up the carcinogen protein, which restores the cellular protein homeostasis, it is likely to alleviate or treat cancers. The PROTAD technology takes advantage of this, and uses the specially designed dual-specific degraders to tag the target proteins as “to be ubiquitined” to achieve targeted degradation. Therefore, the bifunctional compounds exhibit a good inhibitory effect on EGFR, and are good EGFR inhibitors, which can be used to regulate epidermal growth factor receptor (EGFR) and/or its mutants, and suitable for the treatment of receptor tyrosine kinase (RTK)-related diseases, or diseases related to EGFR overexpression or high EGFR activity, wherein the diseases can be specifically selected from tumors, myeloid tumors, or solid tumors, cancers, leukemia, lymphoma, colorectal cancer, brain cancer, bone cancer, epithelial cell-derived tumors (epithelial cancer), basal cell carcinoma, adenocarcinoma, gastrointestinal cancer, lip cancer, oral cancer, esophageal cancer, small intestine cancer, gastric cancer, colon cancer, liver cancer, bladder cancer, pancreatic cancer, ovarian cancer, cervical cancer, lung cancer, breast cancer, skin cancer, squamous cell and/or basal cell carcinoma, prostate cancer, glioma, glioblastoma, renal cell carcinoma and other cancers known to affect systemic epithelial cells, chronic granulocytic leukemia (CML), acute myeloid leukemia (AML), and acute promyelocytic leukemia (APL).
In a third aspect of the present invention, there is provided a pharmaceutical composition comprising the bifunctional compounds provided in the first aspect of the present invention or a pharmaceutically acceptable salt, an isomer, a prodrug, a polymorph, or a solvate thereof, and at least one pharmaceutically acceptable carrier.
In the present invention, the composition may include one or more pharmaceutically acceptable carriers, which generally refer to carriers for administration of therapeutic agents, which themselves do not induce the production of antibodies detrimental to the individual receiving the composition, and do not cause excessive toxicity after administration. These carriers are well known to those skilled in the art, e.g., those described in Remington's Pharmaceutical Sciences (Mack Pub. Co., N. J. 1991). Specifically, the carrier may include, but is not limited to, a combination of one or more of saline, buffer, glucose, water, glycerol, ethanol, adjuvant, etc.
The pharmaceutical composition of the present invention can comprise the bifunctional compound as a sole active ingredient, or a combination of the bifunctional compound with an additional active ingredient to form a combined formulation. The additional active ingredient may be various drugs that can be used to treat tumors, myeloma or solid tumors, and cancers. The amount of the active ingredient in the composition is usually a safe and effective amount, which should be adjustable for those skilled in the art. For example, the dosages of the bifunctional compound and the active ingredient of the pharmaceutical composition usually depend on the weight of the patient, the type of administration, and the condition and severity of the diseases. For example, the dosage of the bifunctional compound as an active ingredient can usually be 1 to 1000 mg/kg/day, 20 to 200 mg/kg/day, 1 to 3 mg/kg/day, 3 to 5 mg/kg/day, 5 to 10 mg/kg/day, 10 to 20 mg/kg/day, 20 to 30 mg/kg/day, 30 to 40 mg/kg/day, 40 to 60 mg/kg/day, 60 to 80 mg/kg/day, 80 to 100 mg/kg/day, 100 to 150 mg/kg/day, 150 to 200 mg/kg/day, 200 to 300 mg/kg/day, 300 to 500 mg/kg/day, or 500 to 1000 mg/kg/day.
The bifunctional compounds of the present invention can be adapted to any route of administration, which can be oral or parenteral administration, e.g., pulmonary administration, nasal administration, rectal administration and/or intravenous injection, and more specifically intradermal, subcutaneous, intramuscular, intraarticular, intraperitoneal, lung, buccal, sublingual, nasal, transdermal, vaginal, oral or parenteral administration. Those skilled in the art can select a suitable formulation form according to the route of administration, e.g., formulations suitable for oral administration which may include, but are not limited to, pills, tablets, masticatory, capsules, granules, drops or syrups; e.g., formulations suitable for parenteral administration which may include, but are not limited to, solutions, suspensions, reconstitutable dry formulations or sprays; e.g., formulations suitable for rectal administration which may include, but are not limited to, suppositories.
In a fourth aspect of the present invention, there is provided a treatment method comprising: administering to an individual a therapeutically effective amount of the bifunctional compounds provided in the first aspect of the present invention or the pharmaceutical composition provided in the third aspect of the present invention.
In the present invention, “individuals” generally include humans and non-human primates, such as mammals, dogs, cats, horses, sheep, pigs, cattle, etc., which can benefit from treatment with the formulations, kits or combined formulations.
In the present invention, “therapeutically effective amount” generally refers to an amount that can achieve the effect of treating the diseases listed above after a proper administration period.
The bifunctional compound of the present invention is a bifunctional compound based on an epidermal growth factor receptor tyrosine kinase inhibitor, which can not only promote the degradation of EGFR protein, but also inhibit the activity of EGFR kinase and have a significant inhibitory effect on the proliferation of EGFR mutation-positive cells.
In the following description, numerous specific embodiments are set forth in order to provide a thorough understanding of the present invention, and those skilled in the art can easily understand other advantages and effects of the present invention from the content disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and various details in this specification can also be modified or changed based on different viewpoints and applications without departing from the spirit of the present invention.
It should be noted that the process equipments or devices not specifically noted in the following embodiments are all conventional equipments or devices in the art.
In addition, unless otherwise specified, it should be understood that one or more method steps mentioned in the present invention do not exclude that there may be additional method steps before and after the combined steps, or additional method steps inserted between these explicitly mentioned steps.
Unless otherwise specified, it should also be understood that the combined connection relationship between one or more equipments/devices mentioned in the present invention does not exclude that additional equipments/devices may also exist before and after the combined equipments/devices, or additional equipments/devices may also be inserted between the two explicitly mentioned equipments/devices. Moreover, unless otherwise specified, the number of each method step is only a convenient tool for identifying each method step, not to limit the sequence of each method step or to limit the scope of the present invention, and the changes or adjustments of the relative relationship of the number of each method step fall within the scope of the present invention without substantial change in the technical content.
The following abbreviations are used throughout the description and examples:
Boc t-butyloxy carbonyl
Con. concentration
DCM dichloromethane
DMF N,N-dimethylformamide
DMSO dimethyl sulfoxide
DIPEA N, N-diisopropylethylamine
EDCI 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide
ESI electrospray ionization
equiv equivalent
EtOH ethanol
HOAT 1-hydroxy-7-azabenzotriazole
HPLC high performance liquid chromatography
HRMS high resolution mass spectrometry
LC-MS liquid chromatography-mass spectrometry
LRMS low resolution mass spectrometry
LC liquid chromatography
Me methyl
MeCN acetonitrile
MeOH methanol
MS mass spectrum
MW microwave
NMM N-methylmorpholine
NMP N-methylpyrrolidone
1H NMR Proton nuclear magnetic resonance
rt room temperature
TFA trifluoroacetic acid
TLC thin layer chromatography
TMS trimethylsilyl
Xantphos; or X-phos 4,5-Bisdiphenylphosphine-9,9-dimethylxanthene
In the present disclosure, the 1H NMR spectra were recorded on a Bruker-500 MHz nuclear magnetic resonance instrument, by using, as a solvent and an internal standard, CD3OD containing 0.1% TMS (1H NMR in CD3OD; δ=3.31 ppm); or using, as a solvent and an internal standard, CDCl3 containing 0.1% TMS (1H NMR in CDCl3; δ=7.26 ppm); or using, as a solvent and an internal standard, DMSO-d6 containing 0.03% TMS (1H NMR in DMSO-d6; δ=2.50 ppm). LRMS spectrum was recorded on an AB Triple 4600 mass spectrometer, HPLC preparation was measured on a SHIMADZU LC-20AP type instrument, and HPLC purity was measured on a SHIMADZU LC-30AP or Waters 1525 type instrument. Unless otherwise specified, all reactions were performed in the air atmosphere. The reactions were tracked by TLC or LC-MS.
Solvents and reagents are processed as follows:
The groups U and V are as shown in Scheme 1.
General Synthesis Method of Intermediates LM (Thio-Pomalidomide with Alkylene Chain-Carboxylic Acid Group Linker):
wherein n=an integer of 1-10, as shown in Scheme 6.
General Synthesis Method of Intermediates LM (Thio-Pomalidomide with PEG Chain-Carboxylic Acid Group Linker):
wherein n=an integer of 1-5, as shown in Scheme 7.
General Synthesis Method of Intermediates LM (Thio-Lenalidomide with Alkylene Chain-Carboxylic Acid Group Linker):
wherein n=an integer of 1-10, as shown in Scheme 8.
General Synthesis Method of Intermediates LM (Thio-Lenalidomide with PEG Chain-Carboxylic Acid Group Linker):
wherein n=an integer of 1-5, as shown in scheme 9.
General Synthesis Method of Intermediates LM (Lenalidomide with Alkylene Chain-Alkynyl Linker):
wherein n=an integer of 1-10, as shown in scheme 10.
General Synthesis Method of Intermediates LM (Thio-Lenalidomide with Alkylene Chain-Amino Linker):
wherein n=an integer of 1-10, as shown in scheme 11.
General Synthesis Method of Intermediates LM (Thio-Lenalidomide with Alkylene Chain-Bromine Linker):
wherein n=an integer of 1-10, as shown in Scheme 12.
General Synthesis Method of Intermediates LM (Pomalidomide with Alkylene Chain-Iodine Linker):
Dacomitinib Derivative A (SIAIS219183) was Prepared According to Scheme 1.
4-bromo-crotonic acid (825 mg, 5 mmol) was dissolved in 4 mL of oxalyl chloride under an argon atmosphere. Then to the mixture was added a drop of DMF to initiate the reaction. The mixture was stirred at room temperature for 2 h, and rotary evaporated under low-temperature to remove the excess oxalyl chloride. A 100 mL clean egg-shaped flask was sequentially charged with N-(3-chloro-4-fluorophenyl)-7-methoxy-6-aminoquinazolin-4-amine (636 mg, 2 mmol), 4 mL THF, and triethylamine (417 mg, 3 mmol) with stirring at room temperature, followed by addition of a solution of the acyl chloride prepared in-situ in 4 mL THE. The reaction mixture was reacted at room temperature for 1 h. After the reaction was complete, the reaction was quenched with water. The mixture was extracted with ethyl acetate. The organic phase was washed with water and saturated brine, dried over anhydrous sodium sulfate, and rotary evaporated to obtain the crude product. The crude product was subjected to C18 reverse phase column chromatography (eluent:MeOH/water) for purification to give the target product SIAIS219176 as a yellow solid (754 mg, yield 81%). 1H NMR (500 MHz, MeOD) δ 9.25 (s, 1H), 8.77 (s, 1H), 7.94 (dd, J=6.6, 2.6 Hz, 1H), 7.69-7.64 (m, 1H), 7.38 (t, J=8.9 Hz, 1H), 7.34 (s, 1H), 7.07-7.01 (m, 1H), 6.69 (d, J=15.3 Hz, 1H), 4.18 (s, 3H), 3.53-3.48 (m, 2H). HRMS (ESI) m/z: calcd for C19H16BrClFN4O2+ [M+H]+, 465.0124; found, 465.0121.
An egg-shaped flask was sequentially charged with SIAIS219176 (200 mg, 0.43 mmol), 4 mL DMF, N-tert-butoxycarbonylpiperazine (160.2 mg, 0.86 mmol), and potassium carbonate (356.6 mg, 1.29 mmol) at room temperature, followed by evacuation and refilling with argon gas and reacting at 40° C. for 2 h. After the reaction was complete as monitored by TLC, the mixture was subject to a reversed phase C18 column chromatography (eluent: MeOH/water) for purification to give a yellow solid which was used directly in the next step. To a solution of the obtained yellow solid dissolved in DCM (6 mL) was added 2 mL CF3COOH. The mixture was stirred at room temperature for 2 h. After the reaction was complete as monitored by LC-MS, the mixture was rotary evaporated to remove most of the CF3COOH, and the pH was adjusted to alkaline with saturated sodium bicarbonate solution. The resulting mixture was extracted with dichloromethane. The organic phase was dried over anhydrous sodium sulfate, and rotary evaporated to obtain the crude product. The crude product was subjected to C18 reverse phase column chromatography (eluent:MeOH/water) for purification to give the target product SIAIS219183 as a yellow solid (178 mg, total yield of two steps 88%). 1H NMR (500 MHz, MeOD) δ 9.24 (s, 1H), 8.75 (s, 1H), 7.93 (dd, J=6.6, 2.6 Hz, 1H), 7.66 (ddd, J=8.9, 4.1, 2.7 Hz, 1H), 7.37 (t, J=8.9 Hz, 1H), 7.33 (s, 1H), 7.03 (dt, J=15.3, 6.2 Hz, 1H), 6.68 (d, J=15.3 Hz, 1H), 4.17 (s, 3H), 3.51-3.48 (m, 2H), 3.36-3.33 (m, 4H), 2.93 (s, 4H). HRMS (ESI) m/z: calcd for C23H25ClFN6O2+ [M+H]+, 471.1706; found, 471.1706.
Referring to Scheme 1, Dacomitinib derivative B was prepared by using a method similar to that of dacomitinib derivative A in Intermediate Example 1. The synthetic and structural characterization data of the intermediate are as follows:
(E)-N-(4-((3-chloro-4-fluorophenyl)amino)-7-methoxyquinazolin-6-yl)-4-(4-(piperazin-1-yl)piperidin-1-yl)but-2-enamide (SIAIS262021). (yellow solid, 166.3 mg, total yield of two steps 79%)1H NMR (500 MHz, MeOD) δ 9.26 (s, 1H), 8.75 (s, 1H), 7.93 (dd, J=6.6, 2.6 Hz, 1H), 7.67-7.63 (m, 1H), 7.40-7.34 (m, 2H), 7.05-7.01 (m, 1H), 6.84 (d, J=15.2 Hz, 1H), 4.17 (s, 3H), 4.03 (d, J=7.0 Hz, 2H), 3.66 (s, 1H), 3.29-3.26 (m, 4H), 3.22-3.09 (m, 2H), 3.01-2.80 (m, 6H), 2.16 (s, 2H), 2.01-1.85 (m, 2H). HRMS (ESI) m/z: calcd for C28H34ClFN7O2+ [M+H]+, 554.2441; found, 554.2433.
6-acetoxy-7-methoxy-3,4-dihydroquinazolin-4(3H)-one (468.4 mg, 2 mmol) was dissolved in 4 mL of sulfoxide chloride under an argon atmosphere. Then to the mixture was added a drop of DMF to initiate the reaction. The mixture was stirred at 90° C. for 4 h, and rotary evaporated under low-temperature to remove the excess sulfoxide chloride, and the obtained residue was used directly in the next step. A 100 mL clean egg-shaped flask was sequentially charged with the product obtained from the previous step (2 mmol), 10 mL DMF, and 3,4-dichloro-2-fluoroaniline (432 mg, 2.4 mmol) with stirring at 80° C. for 1 h. After the reaction was complete, the reaction was quenched with ice water, and the mixture was filtered to obtain the intermediate which was used directly in the next step. A 100 mL clean egg-shaped flask was sequentially charged with the intermediate obtained from the previous step (2 mmol), 10 mL methanol, and 2 mL aqueous ammonia solution with stirring under reflux at 70° C. for 2 h. The reaction mixture was cooled, filtered, and washed with a small amount of cold methanol to give the target product SIAIS219148 as a yellow solid (310 mg, total yield of three steps 44%). 1H NMR (500 MHz, MeOD) δ 8.38 (s, 1H), 7.78 (s, 1H), 7.61 (t, J=8.2 Hz, 1H), 7.40 (dd, J=8.8, 1.9 Hz, 1H), 7.21 (s, 1H). HRMS (ESI) m/z: calcd for C15H11Cl2FN3O2+ [M+H]+, 354.0207; found, 354.0202.
An egg-shaped flask was sequentially charged with SIAIS219148 (177.1 mg, 0.5 mmol), 4 mL DMF, tert-butyl 4-(tosyloxy)piperidine-1-carboxylate (213.3 mg, 0.6 mmol), and potassium carbonate (207.3 mg, 1.5 mmol) at room temperature, followed by evacuation and refilling with argon gas and reacting at 70° C. for 5 h. After the reaction was complete as monitored by TLC, the mixture was subject to a reversed phase C18 column chromatography (eluent: MeOH/water) for purification to give a yellow solid which was used directly in the next step. To a solution of the obtained yellow solid dissolved in DCM (6 mL) was added 2 mL CF3COOH. The mixture was stirred at room temperature for 2 h. After the reaction was complete as monitored by LC-MS, the mixture was rotary evaporated to remove most of the CF3COOH, and the pH was adjusted to alkaline with saturated sodium bicarbonate solution. The resulting mixture was extracted with dichloromethane. The organic phase was dried over anhydrous sodium sulfate, and rotary evaporated to obtain the crude product. The crude product was subjected to C18 reverse phase column chromatography (eluent: MeOH/water) for purification to give the target product SIAIS219149B as a yellow solid (196.8 mg, total yield of two steps 90%). 1H NMR (500 MHz, MeOD) δ 8.37 (s, 1H), 7.79 (s, 1H), 7.59 (t, J=8.2 Hz, 1H), 7.42 (dd, J=8.8, 1.9 Hz, 1H), 7.20 (s, 1H), 4.85-4.80 (m, 1H), 4.00 (s, 3H), 3.98-3.89 (m, 2H), 3.71-3.59 (m, 2H), 2.08 (dd, J=8.4, 3.7 Hz, 2H), 1.90 (dd, J=10.1, 6.6 Hz, 2H). HRMS (ESI) m/z: calcd for C20H20Cl2FN4O2+ [M+H]+, 437.0942; found, 437.0942.
4-((3-chloro-4-fluorophenyl)amino)-7-methoxyquinazolin-6-ol (639.4 mg, 2 mmol), 5 mL DMF, 1-bromo-3-chloropropane (630 mg, 4 mmol), and potassium carbonate (829.3 mg, 6 mmol) were reacted at room temperature under an argon atmosphere for 12 h. After the reaction was complete, the mixture was subjected to C18 reverse phase column chromatography (eluent:MeOH/water) for separation to give the target product SIAIS184151 as a yellow solid (350 mg, yield 44%). 1H NMR (500 MHz, MeOD) δ 8.44 (s, 1H), 8.00 (dd, J=6.7, 2.6 Hz, 1H), 7.75 (s, 1H), 7.67 (ddd, J=8.8, 4.0, 2.7 Hz, 1H), 7.25 (t, J=9.0 Hz, 1H), 7.17 (s, 1H), 4.33 (t, J=5.9 Hz, 2H), 4.00 (s, 3H), 3.84 (t, J=6.3 Hz, 2H), 2.34 (p, J=6.1 Hz, 2H). HRMS (ESI) m/z: calcd for C18H17Cl2FN3O2+ [M+H]+, 396.0676; found, 396.0676.
An egg-shaped flask was sequentially charged with SIAIS184151 (350 mg, 0.883 mmol), 4 mL NMP, N-tert-butoxycarbonylpiperazine (328.9 mg, 1.766 mmol), DIPEA (456.5 mg, 3.532 mmol), and NaI (264.7 mg, 1.766 mmol) at room temperature, followed by evacuation and refilling with argon gas and reacting at 90° C. for 2 h. After the reaction was complete as monitored by TLC, the mixture was subject to a reversed phase C18 column chromatography (eluent: MeOH/water) for separation to give a yellow solid which was used directly in the next step. To a solution of the obtained yellow solid dissolved in DCM (6 mL) was added 2 mL CF3COOH. The mixture was stirred at room temperature for 2 h. After the reaction was complete as monitored by LC-MS, the mixture was rotary evaporated to remove most of the CF3COOH, and the pH was adjusted to alkaline with saturated sodium bicarbonate solution. The resulting mixture was extracted with dichloromethane, dried over anhydrous sodium sulfate, and rotary evaporated to obtain the crude product. The crude product was subjected to C18 reverse phase column chromatography (eluent: MeOH/water) for separation to give the target product SIAIS184161 as a yellow solid (320 mg, total yield of two steps 81%). 1H NMR (500 MHz, MeOD) δ 8.74 (s, 1H), 7.98 (s, 1H), 7.94 (dd, J=6.6, 2.6 Hz, 1H), 7.69-7.63 (m, 1H), 7.37 (t, J=8.9 Hz, 1H), 7.27 (s, 1H), 4.37 (t, J=5.6 Hz, 2H), 4.09 (s, 3H), 3.57-3.52 (m, 4H), 3.48-3.40 (m, 4H), 3.32 (d, J=4.4 Hz, 2H), 2.41-2.32 (m, 2H). HRMS (ESI) m/z: calcd for C22H26ClFN5O2+ [M+H]+, 446.1754; found, 446.1754.
Referring to Scheme 3, Gefitinib derivative B was prepared by using a method similar to that of Gefitinib derivative A in Intermediate Example 4. The synthetic and structural characterization data of the intermediate are as follows:
N-(3-chloro-4-fluorophenyl)-7-methoxy-6-(3-(4-(piperazin-1-yl)piperidin-1-yl)propoxy)quinazolin-4-amine (SIAIS262080). (yellow solid, 401 mg, total yield of two steps 76%)1H NMR (500 MHz, MeOD) δ 8.75 (s, 1H), 7.99 (s, 1H), 7.94 (dd, J=6.6, 2.6 Hz, 1H), 7.71-7.62 (m, 1H), 7.38 (t, J=8.9 Hz, 1H), 7.29 (s, 1H), 4.38 (t, J=5.6 Hz, 2H), 4.08 (s, 3H), 3.58-3.52 (m, 8H), 3.49-3.40 (m, 9H), 3.34 (d, J=4.4 Hz, 2H), 2.41-2.31 (m, 2H). HRMS (ESI) m/z: calcd for C27H35ClFN6O2+ [M+H]+, 529.2489, found, 529.2489.
Canertinib derivative A (SIAIS293064) was prepared according to Scheme 4.
To a solution of tert-butyl 4-(3-((6-amino-4-((3-chloro-4-fluorophenyl)amino)quinazolin-7-yl)oxy)propyl)piperazine-1-carboxylat e (531 mg, 1 mmol) in 5 mL of THF was added acryloyl chloride (362 mg, 4 mmol) in 0° C. ice-water bath under an argon atmosphere. The mixture was then warmed up to room temperature and reacted for 1 h. After the reaction was complete, the reaction was quenched with water. The mixture was extracted with dichloromethane. The organic phase was dried over anhydrous sodium sulfate, and rotary evaporated to obtain the intermediate which was then subjected to silica gel column chromatography (eluent (v/v): dichloromethane/methanol=20:1 to 10:1) for purification to give a yellow solid which was used directly in the next step. To a solution of the obtained yellow solid dissolved in DCM (6 mL) was added 2 mL CF3COOH. The mixture was stirred at room temperature for 2 h. After the reaction was complete as monitored by LC-MS, the mixture was rotary evaporated to remove most of the CF3COOH, and the pH was adjusted to alkaline with saturated sodium bicarbonate solution. The resulting mixture was extracted with dichloromethane. The organic phase was dried over anhydrous sodium sulfate, and rotary evaporated to obtain the crude product. The crude product was subjected to C18 reverse phase column chromatography (eluent:MeOH/water) for separation to give the target product SIAIS293064 as a yellow solid (341 mg, total yield of two steps 70%). 1H NMR (500 MHz, MeOD) δ 9.20 (s, 1H), 8.76 (s, 1H), 7.95-7.91 (m, 1H), 7.66 (ddd, J=8.9, 4.2, 2.6 Hz, 1H), 7.37 (d, J=5.5 Hz, 1H), 6.89 (dd, J=16.9, 10.3 Hz, 1H), 6.52 (dd, J=16.9, 1.5 Hz, 1H), 5.90 (dd, J=10.3, 1.6 Hz, 1H), 4.51 (t, J=5.8 Hz, 2H), 3.72 (dd, J=21.9, 11.5 Hz, 6H), 3.61-3.56 (m, 2H), 2.54 (td, J=11.7, 5.8 Hz, 2H). HRMS (ESI) m/z: calcd for C24H27ClFN6O2+ [M+H]+, 485.1863; found, 485.1861.
Referring to Scheme 4, Canertinib Derivative B was prepared by using a method similar to that of Canertinib Derivative A in Intermediate Example 6. The synthetic and structural characterization data of the intermediate are as follows:
N-(4-((3-chloro-4-fluorophenyl)amino)-7-(3-(4-(piperazin-1-yl)piperidin-1-yl)propoxy)quinazolin-6-yl)acrylamide (SIAIS249183). (yellow solid, 200 mg, total yield of two steps 62%)1H NMR (500 MHz, MeOD) δ 9.22 (s, 1H), 8.78 (s, 1H), 7.96-7.91 (m, 1H), 7.67-7.62 (m, 1H), 7.38 (d, J=5.5 Hz, 1H), 6.89-6.83 (m, 1H), 6.55-6.51 (m, 1H), 5.91 (dd, J=10.3, 1.6 Hz, 1H), 4.52 (t, J=5.8 Hz, 2H), 3.79-3.68 (m, 10H), 3.64-3.56 (m, 3H), 2.54-2.43 (m, 6H). HRMS (ESI) m/z: calcd for C29H36ClFN7O2+ [M+H]+, 568.2598, found, 568.2591.
Gefitinib derivative C (SIAIS293033) was prepared according to Scheme 5.
To a solution of N-(3-chloro-4-fluorophenyl)-7-methoxy-6-(piperidin-4-yloxy)quinazolin-4-amine (400 mg, 0.8 mmol) in 5 mL NMP were sequentially added 3-(tert-butoxy)-3-oxopropanoic acid (234 mg, 1.2 mmol), NaI (240 mg, 1.6 mmol), and DIPEA (310 mg, 2.4 mmol) under air atmosphere. The mixture was reacted at 80° C. for 2 h. After the reaction was complete, the reaction was quenched with water. The mixture was extracted with dichloromethane. The organic phase was dried over anhydrous sodium sulfate, and rotary evaporated to obtain the intermediate which was then subjected to silica gel column chromatography (eluent (v/v): dichloromethane/methanol=20:1 to 10:1) for purification to give a yellow solid which was used directly in the next step. To a solution of the obtained yellow solid dissolved in DCM (6 mL) was added 2 mL CF3COOH. The mixture was reacted at room temperature for 2 h. After the reaction was complete as monitored by LC-MS, the mixture was rotary evaporated to remove most of the CF3COOH, and the pH was adjusted to alkaline with saturated sodium bicarbonate solution. The resulting mixture was extracted with dichloromethane. The organic phase was dried over anhydrous sodium sulfate, and rotary evaporated to obtain the crude product. The crude product was subjected to C18 reverse phase column chromatography (eluent: MeOH/water) for purification to give the target product SIAIS293033 as a yellow solid (320 mg, total yield of two steps 87%). 1H NMR (500 MHz, MeOD) δ 8.74 (s, 1H), 8.11 (s, 1H), 7.93 (d, J=4.5 Hz, 1H), 7.66 (d, J=8.8 Hz, 1H), 7.37 (t, J=8.9 Hz, 1H), 7.30 (s, 1H), 4.95 (s, 2H), 4.12 (s, 1H), 4.10 (s, 3H), 3.57 (d, J=15.2 Hz, 4H), 2.34 (s, 4H). HRMS (ESI) m/z: calcd for C22H23ClFN4O4+ [M+H]+, 461.1386, found, 461.1385.
Referring to Scheme 5, Sapitinib Derivative A was prepared by using a method similar to that of Gefitinib derivative C in Intermediate Example 8. The synthetic and structural characterization data of the intermediate are as follows:
2-(4-((4-((3-chloro-2-fluorophenyl)amino)-7-methoxyquinazolin-6-yl)oxy)piperidin-1-yl)acetic acid (SIAIS293061). (yellow solid, 335 mg, total yield of two steps 61%) 1H NMR (500 MHz, MeOD) δ 8.67 (s, 1H), 8.21 (s, 1H), 7.62 (d, J=7.6 Hz, 2H), 7.53-7.48 (m, 3H), 7.33-7.27 (m, 2H), 4.94 (s, 2H), 4.14 (s, 1H), 4.11 (s, 3H), 3.58 (d, J=15.2 Hz, 4H), 2.34 (s, 4H). HRMS (ESI) m/z: calcd for C22H23ClFN4O4+ [M+H]+, 461.1386, found, 461.1381.
The synthetic methods and structural characterization data of the intermediates SIAIS151001, SIAIS151004, SIAIS151005, SIAIS151006, SIAIS151007, SIAIS151025, SIAIS151026, SIAIS151019, SIAIS151020, SIAIS151027, SIAIS151086, SIAIS1204057, SIAIS1204085, SIAIS1210133, SIAIS1204061, SIAIS1210133, SIAIS1204061, SIAIS120, SIA151074, SIAIS0101061, SIAIS120, SIAIS151008 SIAIS074012, SIAIS074013, SIAIS074014, SIAIS074015, SIAIS074016, SIAIS074019, SIAIS074020, and SIAIS172147 that act as linkers, can be seen in China Appl. Pub. No. CN109912655 A.
The synthetic methods and structural characterization data of the intermediates SIAIS1220099, SIAIS299138, SIAIS299135, SIAIS213132, SIAIS213135, SIAIS1216135, SIAIS1216137, SIAIS1220059, SIAIS1220013, SIAIS1220015, and SIAIS1220141 that act as linkers, can be seen in China Pat. No. 201910279248.9.
The synthetic methods and structural characterization data of the intermediates SIAIS164118 and SIAIS164119 that act as linkers, can be seen in PCT publication WO 2019170150 (A1).
The synthetic methods and structural characterization data of the intermediates SIAIS1213061 and SIAIS1213011 that act as linkers, can be seen in PCT publication WO2020103878 (A1).
According to Scheme 7, a 50 mL egg-shaped flask was sequentially charged with the intermediate compound SIAIS151014 (0.724 mmol, 1 equiv), anhydrous N,N-dimethylformamide (10 mL) and anhydrous potassium carbonate (1.448 mmol, 2 equiv), followed by slow addition of the corresponding p-toluenesulfonate-substituted substrate (0.869 mmol, 1.2 equiv) as a linker with stirring at room temperature. After the completion of addition, the mixture was stirred at room temperature for 0.5 h. After the starting materials were consumed, the reaction mixture was filtered to remove the insoluble substance, and the filtrate was then directly subjected to a reverse phase C18 column chromatography (eluent (v/v): acetonitrile/water=10%-100%) for separation, and the collected fractions were concentrated under reduced pressure to remove the solvents, to give the corresponding tert-butyl ester intermediate compound. The corresponding tert-butyl ester intermediate compound, dichloromethane (1 mL) and trifluoroacetic acid (3 mL) were sequentially added to a 25 mL egg-shaped flask, and stirred at room temperature for 1 h, and then concentrated under reduced pressure to remove the solvents. The resulting residue was treated by addition of water and lyophilized to afford the corresponding target compound SIAIS1204137 (light yellow solid, 185 mg, yield 69%). 1H NMR (500 MHz, DMSO) δ 11.12 (s, 1H), 7.83-7.73 (m, 2H), 7.64 (d, J=6.6 Hz, 1H), 5.12 (dd, J=12.8, 5.4 Hz, 1H), 4.08 (s, 2H), 3.77 (t, J=6.4 Hz, 2H), 3.14-3.07 (m, 2H), 2.94-2.82 (m, 1H), 2.66-2.55 (m, 2H), 2.09-2.01 (m, 1H). HRMS (ESI) m/z: calcd for C17H17N2O7S+ [M+H]+, 393.0751; found, 393.0763.
The compound SIAIS1204139 was prepared according to the method of intermediate example 10, except that the p-toluenesulfonate-substituted substrate was tert-butyl 2-(2-(2-(tosyloxy)ethoxy)ethoxy)acetate. The target compound SIAIS1204139 was obtained as a light yellow solid (190 mg, yield 63%). 1H NMR (500 MHz, DMSO) δ 11.12 (s, 1H), 7.83-7.76 (m, 2H), 7.63 (dd, J=6.4, 1.3 Hz, 1H), 5.12 (dd, J=12.9, 5.4 Hz, 1H), 4.02 (s, 2H), 3.72 (t, J=6.3 Hz, 2H), 3.59 (s, 4H), 3.39-3.30 (m, 2H), 3.13-3.06 (m, 1H), 2.64-2.52 (m, 2H), 2.09-2.02 (m, 1H). HRMS (ESI) m/z: calcd for C19H21BN2O8S+ [M+H]+, 437.1013; found, 437.1032.
The compound SIAIS1204141 was prepared according to the method of intermediate example 10, except that the p-toluenesulfonate-substituted substrate was tert-butyl 2-(2-(2-(2-(tosyloxy)ethoxy)ethoxy)ethoxy)acetate. The target compound SIAIS1204141 was obtained as a light yellow solid (246 mg, yield 74%). 1H NMR (500 MHz, DMSO) δ 11.12 (s, 1H), 7.85-7.73 (m, 2H), 7.63 (dd, J=6.1, 1.9 Hz, 1H), 5.12 (dd, J=12.9, 5.4 Hz, 1H), 4.02 (s, 2H), 3.71 (t, J=6.3 Hz, 2H), 3.62-3.48 (m, 8H), 3.35 (t, J=6.3 Hz, 2H), 2.94-2.84 (m, 1H), 2.63-2.52 (m, 2H), 2.11-1.99 (m, 1H). HRMS (ESI) m/z: calcd for C21H25N2O9S+ [M+H]+, 481.1275; found, 481.1273.
The compound SIAIS1204147 was prepared according to the method of intermediate example 10, except that the p-toluenesulfonate-substituted substrate was tert-butyl 14-(tosyloxy)-3,6,9,12-tetraoxatetradecanoate. The target compound SIAIS1204147 was obtained as a light yellow solid (228 mg, yield 63%). 1H NMR (500 MHz, DMSO) δ 11.12 (s, 1H), 7.83-7.73 (m, 2H), 7.63 (dd, J=6.2, 1.7 Hz, 1H), 5.12 (dd, J=12.9, 5.4 Hz, 1H), 4.01 (s, 2H), 3.71 (t, J=6.3 Hz, 2H), 3.59-3.54 (m, 4H), 3.54-3.49 (m, 8H), 3.35 (t, J=6.3 Hz, 2H), 2.94-2.84 (m, 1H), 2.64-2.56 (m, 1H), 2.55-2.51 (m, 1H), 2.08-2.02 (m, 1H). HRMS (ESI) m/z: calcd for C23H29N2O10S+ [M+H]+, 525.1537; found, 525.1536.
The compound SIAIS1204149 was prepared according to the method of intermediate example 10, except that the p-toluenesulfonate-substituted substrate was tert-butyl 17-(tosyloxy)-3,6,9,12,15-pentaoxaheptadecanoate. The target compound SIAIS1204149 was obtained as a light yellow solid (259 mg, yield 66%). 1H NMR (500 MHz, DMSO) δ 11.12 (s, 1H), 7.83-7.74 (m, 2H), 7.63 (dd, J=6.2, 1.8 Hz, 1H), 5.12 (dd, J=12.9, 5.4 Hz, 1H), 4.01 (s, 2H), 3.71 (t, J=6.3 Hz, 2H), 3.60-3.55 (m, 4H), 3.55-3.47 (m, 12H), 3.35 (t, J=6.3 Hz, 2H), 2.93-2.84 (m, 1H), 2.64-2.56 (m, 1H), 2.55-2.51 (m, 1H), 2.08-2.02 (m, 1H). HRMS (ESI) m/z: calcd for C25H33N2O11S+ [M+H]+, 569.1800; found, 569.1814.
The compound SIAIS151045 was prepared according to the method of Scheme 6, except that the brominated substrate as the linker was tert-butyl 2-bromoacetate. The target compound SIAIS151045 was obtained as a light yellow solid (0.69 g, yield 80%). 1H NMR (500 MHz, DMSO) δ 13.06 (s, 1H), 11.15 (s, 1H), 7.80 (dd, J=8.1, 7.3 Hz, 1H), 7.66 (t, J=7.9 Hz, 2H), 5.13 (dd, J=12.9, 5.4 Hz, 1H), 4.09 (s, 2H), 2.92-2.85 (m, 1H), 2.66-2.51 (m, 2H), 2.08-2.03 (m, 1H). HRMS (ESI) m/z: calcd for C15H13N2O6S+ [M+H]+, 349.0489; found, 349.0297.
The compound SIAIS151138B was prepared according to the method of intermediate example 15, except that the brominated substrate as the linker was tert-butyl 3-bromopropionate. The target compound SIAIS151138B was obtained as a light yellow solid (0.64 g, yield 74%). 1H NMR (500 MHz, DMSO) δ 11.12 (s, 1H), 7.81-7.76 (m, 2H), 7.64 (d, J=6.7 Hz, 1H), 5.11 (dd, J=12.9, 5.4 Hz, 1H), 3.32 (t, J=7.0 Hz, 2H), 2.92-2.84 (m, 1H), 2.66 (t, J=7.0 Hz, 2H), 2.62-2.51 (m, 2H), 2.07-2.00 (m, 1H). HRMS (ESI) m/z: calcd for C16H15N2O6S+ [M+H]+, 363.0645; found, 363.0802.
The compound SIAIS151139B was prepared according to the method of intermediate example 15, except that the brominated substrate as the linker was tert-butyl 4-bromobutyrate. The target compound SIAIS151139B was obtained as a light yellow solid (0.71 g, yield 82%). 1H NMR (500 MHz, DMSO) δ 12.24 (s, 1H), 11.12 (s, 1H), 7.86-7.74 (m, 2H), 7.63 (d, J=6.2 Hz, 1H), 5.11 (dd, J=12.9, 5.4 Hz, 1H), 3.15 (t, J=7.2 Hz, 2H), 2.92-2.84 (m, 1H), 2.64-2.51 (m, 2H), 2.42 (t, J=7.2 Hz, 2H), 2.09-2.02 (m, 1H), 1.93-1.83 (m, 2H). HRMS (ESI) m/z: calcd for C17H17N2O6S+ [M+H]+, 377.0802; found, 377.0962.
The compound SIAIS151140B was prepared according to the method of intermediate example 15, except that the brominated substrate as the linker was tert-butyl 5-bromopentanoate. The target compound SIAIS151140B was obtained as a light yellow solid (0.9 g, yield 74%). 1H NMR (500 MHz, DMSO) δ 11.12 (s, 1H), 7.83-7.71 (m, 2H), 7.62 (d, J=6.9 Hz, 1H), 5.11 (dd, J=12.9, 5.4 Hz, 1H), 3.13 (t, J=6.6 Hz, 2H), 2.92-2.85 (m, 1H), 2.64-2.52 (m, 2H), 2.28 (t, J=6.6 Hz, 2H), 2.08-2.02 (m, 1H), 1.72-1.65 (m, 4H). HRMS (ESI) m/z: calcd for C18H19N2O6S+ [M+H]+, 391.0958; found, 391.1109.
The compound SIAIS151141B was prepared according to the method of intermediate example 15, except that the brominated substrate as the linker was tert-butyl 6-bromohexanoate. The target compound SIAIS151141B was obtained as a light yellow solid (0.71 g, yield 74%). 1H NMR (500 MHz, DMSO) δ 12.01 (s, 1H), 11.12 (s, 1H), 7.82-7.70 (m, 2H), 7.62 (d, J=7.1 Hz, 1H), 5.11 (dd, J=12.9, 5.4 Hz, 1H), 3.12 (t, J=7.2 Hz, 2H), 2.92-2.85 (m, 1H), 2.62-2.48 (m, 2H), 2.22 (t, J=7.2 Hz, 2H), 2.08-2.03 (m, 1H), 1.71-1.63 (m, 2H), 1.59-1.51 (m, 2H), 1.49-1.40 (m, 2H). HRMS (ESI) m/z: calcd for C19H21N2O6S+ [M+H]+, 405.1115; found, 405.1268.
The compound SIAIS151142B was prepared according to the method of intermediate example 15, except that the brominated substrate as the linker was tert-butyl 7-bromoheptanoate. The target compound SIAIS151142B was obtained as a light yellow solid (0.7 g, yield 80%). 1H NMR (500 MHz, DMSO) δ 11.12 (s, 1H), 7.80-7.71 (m, 2H), 7.62 (d, J=6.9 Hz, 1H), 5.11 (dd, J=12.9, 5.4 Hz, 1H), 3.12 (t, J=7.3 Hz, 2H), 2.92-2.85 (m, 1H), 2.62-2.52 (m, 2H), 2.20 (t, J=7.3 Hz, 2H), 2.07-2.00 (m, 1H), 1.69-1.62 (m, 2H), 1.53-1.47 (m, 2H), 1.46-1.41 (m, 2H), 1.36-1.27 (m, 2H). HRMS (ESI) m/z: calcd for C20H23N2O6S+ [M+H]+, 419.1271; found, 419.1432.
According to Scheme 9, a 50 mL egg-shaped flask was sequentially charged with the intermediate compound SIAIS171095 (0.724 mmol, 1 equiv), anhydrous N,N-dimethylformamide (10 mL) and anhydrous potassium carbonate (1.448 mmol, 2 equiv), followed by slow addition of the corresponding p-toluenesulfonate-substituted substrate (0.869 mmol, 1.2 equiv) as a linker with stirring at room temperature. After the completion of addition, the mixture was stirred at room temperature for 0.5 h. After the starting materials were consumed, the reaction mixture was filtered to remove the insoluble substance, and the filtrate was then directly subjected to a reverse phase C18 column chromatography (eluent (v/v): acetonitrile/water=10%-100%) for separation, and the collected fractions were concentrated under reduced pressure to remove the solvents, to give the corresponding tert-butyl ester intermediate compound. The corresponding tert-butyl ester intermediate compound, dichloromethane (1 mL) and trifluoroacetic acid (3 mL) were sequentially added to a 25 mL egg-shaped flask, and stirred at room temperature for 1 h, and then concentrated under reduced pressure to remove the solvents. The resulting residue was treated by addition of water and lyophilized to afford the corresponding target compound SIAIS1213129 (light yellow solid, 148 mg, yield 54%). 1H NMR (500 MHz, CDCl3) δ 8.90 (s, 1H), 7.81 (d, J=7.5 Hz, 1H), 7.68 (d, J=7.7 Hz, 1H), 7.54 (t, J=7.7 Hz, 1H), 5.33 (dd, J=13.4, 5.1 Hz, 1H), 4.60 (d, J=17.2 Hz, 1H), 4.47 (d, J=17.2 Hz, 1H), 4.11 (s, 2H), 3.78-3.73 (m, 1H), 3.72-3.66 (m, 1H), 3.22 (t, J=6.2 Hz, 2H), 2.98-2.93 (m, 1H), 2.90-2.82 (m, 1H), 2.53-2.43 (m, 1H), 2.32-2.25 (m, 1H). HRMS (ESI) m/z: calcd for C17H19N2O6S+ [M+H]+, 379.0958; found, 379.0963.
The compound SIAIS1213131 was prepared according to the method of intermediate example 21, except that the p-toluenesulfonate-substituted substrate was tert-butyl 2-(2-(2-(tosyloxy)ethoxy)ethoxy)acetate. The target compound SIAIS1213131 was obtained as a light yellow solid (158 mg, yield 52%). 1H NMR (500 MHz, CDCl3) δ 8.77 (s, 1H), 7.68 (d, J=7.5 Hz, 1H), 7.53 (d, J=7.7 Hz, 1H), 7.42 (t, J=7.7 Hz, 1H), 5.21 (dd, J=13.4, 5.1 Hz, 1H), 4.41 (d, J=17.1 Hz, 1H), 4.32 (d, J=17.1 Hz, 1H), 4.06 (s, 2H), 3.65-3.59 (m, 4H), 3.54 (t, J=4.1 Hz, 2H), 3.11 (t, J=6.1 Hz, 2H), 2.88-2.83 (m, 1H), 2.81-2.76 (m, 1H), 2.42-2.34 (m, 1H), 2.20-2.14 (m, 1H). HRMS (ESI) m/z: calcd for C19H23BN2O7S+ [M+H]+, 423.1200; found, 423.1205.
The compound SIAIS1213133 was prepared according to the method of intermediate example 21, except that the p-toluenesulfonate-substituted substrate was tert-butyl 2-(2-(2-(2-(tosyloxy)ethoxy)ethoxy)ethoxy)acetate. The target compound SIAIS1213133 was obtained as a light yellow oil (149 mg, yield 44%). 1H NMR (500 MHz, CDCl3) δ 8.91 (s, 1H), 7.75 (d, J=7.5 Hz, 1H), 7.61 (d, J=7.6 Hz, 1H), 7.50 (t, J=7.7 Hz, 1H), 5.29 (dd, J=13.4, 5.1 Hz, 1H), 4.49 (d, J=17.0 Hz, 1H), 4.39 (d, J=17.1 Hz, 1H), 4.17-4.15 (m, 2H), 3.72-3.63 (m, 10H), 3.20 (t, J=6.3 Hz, 2H), 2.96-2.90 (m, 1H), 2.90-2.82 (m, 1H), 2.50-2.44 (m, 1H), 2.28-2.22 (m, 1H). HRMS (ESI) m/z: calcd for C21H27N2O8S+ [M+H]+, 467.1483; found, 467.1467.
The compound SIAIS1213135 was prepared according to the method of intermediate example 21, except that the p-toluenesulfonate-substituted substrate was tert-butyl 14-(tosyloxy)-3,6,9,12-tetraoxatetradecanoate. The target compound SIAIS1213135 was obtained as a light yellow oil (181 mg, yield 49%). 1H NMR (500 MHz, CDCl3) δ 8.61 (s, 1H), 7.78 (dd, J=7.6, 0.7 Hz, 1H), 7.63 (dd, J=7.8, 0.8 Hz, 1H), 7.50 (t, J=7.0 Hz, 1H), 5.29 (dd, J=13.3, 5.1 Hz, 1H), 4.50 (d, J=17.0 Hz, 1H), 4.40 (d, J=16.9 Hz, 1H), 4.15 (s, 2H), 3.72-3.66 (m, 14H), 3.19 (t, J=6.6 Hz, 2H), 2.95-2.93 (m, 1H), 2.91-2.86 (m, 1H), 2.52-2.46 (m, 1H), 2.28-2.24 (m, 1H). HRMS (ESI) m/z: calcd for C23H31N2O9S+ [M+H]+, 511.1745; found, 511.1749.
The compound SIAIS1213133 was prepared according to the method of intermediate example 21, except that the p-toluenesulfonate-substituted substrate was tert-butyl 17-(tosyloxy)-3,6,9,12,15-pentaoxaheptadecanoate. The target compound SIAIS1213133 was obtained as a light yellow oil (209 mg, yield 52%). 1H NMR (500 MHz, CDCl3) δ 8.71 (s, 1H), 7.77 (d, J=7.0 Hz, 1H), 7.64 (dd, J=7.7, 0.7 Hz, 1H), 7.54-7.49 (m, 1H), 5.31 (dd, J=13.4, 5.1 Hz, 1H), 4.50 (d, J=17.0 Hz, 1H), 4.40 (d, J=17.0 Hz, 1H), 4.17 (s, 2H), 3.76-3.74 (m, 2H), 3.70-3.66 (m, 12H), 3.64-3.61 (m, 4H), 3.20 (t, J=6.5 Hz, 2H), 2.98-2.94 (m, 1H), 2.90-2.85 (m, 1H), 2.53-2.43 (m, 1H), 2.30-2.25 (m, 1H). HRMS (ESI) m/z: calcd for C25H35N2O10S+ [M+H]+, 569.1800; found, 569.1814.
The compound SIAIS171090 was prepared according to the method of Scheme 8, except that the brominated substrate as the linker was tert-butyl 2-bromoacetate. The target compound SIAIS171090 was obtained as a white solid (77 mg, total yield of step 3: 64%). 1H NMR (500 MHz, DMSO) δ 12.88 (s, 1H), 11.00 (s, 1H), 7.68-7.45 (m, 3H), 5.15-5.13 (m, 1H), 4.32 (dd, J=56.2, 17.3 Hz, 2H), 3.94 (s, 2H), 2.95-2.91 (m, 1H), 2.63-2.59 (m, 1H), 2.49-2.39 (m, 1H), 2.08-1.92 (m, 1H). HRMS (ESI) m/z: calcd for C15H15N2O5S+ [M+H]+, 335.0696; found, 334.8134.
The compound SIAIS171086 was prepared according to the method of intermediate example 26, except that the brominated substrate as the linker was tert-butyl 3-bromopropionate. The target compound SIAIS171086 was obtained as a white solid (40 mg, total yield of step 3: 32%). 1H NMR (500 MHz, DMSO) δ 10.99 (s, 1H), 7.70-7.55 (m, 3H), 5.13 (dd, J=13.3, 5.1 Hz, 1H), 4.40-4.18 (m, 2H), 3.24 (t, J=7.0 Hz, 2H), 2.95-2.87 (m, 1H), 2.63-2.53 (m, 3H), 2.47-2.34 (m, 1H), 2.05-1.95 (m, 1H). HRMS (ESI) m/z: calcd for C16H17N2O5S+ [M+H]+, 349.0853; found, 348.8166.
The compound SIAIS171089 was prepared according to the method of intermediate example 26, except that the brominated substrate as the linker was tert-butyl 4-bromobutyrate. The target compound SIAIS171089 was obtained as a white solid (50 mg, total yield of step 3: 38%). 1H NMR (500 MHz, DMSO) δ 12.15 (s, 1H), 10.99 (s, 1H), 7.71-7.49 (m, 3H), 5.13 (dd, J=13.3, 5.1 Hz, 1H), 4.41-4.18 (m, 2H), 3.10 (t, J=7.3 Hz, 2H), 2.92-2.88 (m, 1H), 2.61-2.59 (m, 1H), 2.49-2.42 (m, 1H), 2.38 (t, J=7.2 Hz, 2H), 2.05-1.96 (m, 1H), 1.84-1.74 (m, 2H). HRMS (ESI) m/z: calcd for C17H19N2O5S+ [M+H]+, 363.1009; found, 362.8160.
The compound SIAIS171079 was prepared according to the method of intermediate example 26, except that the brominated substrate as the linker was tert-butyl 5-bromopentanoate. The target compound SIAIS171079 was obtained as a white solid (30 mg, total yield of step 3: 22%). 1H NMR (500 MHz, DMSO) δ 12.01 (s, 1H), 10.98 (s, 1H), 7.66-7.55 (m, 3H), 5.12 (dd, J=13.3, 5.1 Hz, 1H), 4.37-4.18 (m, 2H), 3.10-3.05 (m, 2H), 2.95-2.84 (m, 1H), 2.65-2.61 (m, 1H), 2.48-2.38 (m, 1H), 2.27-2.20 (m, 3H), 1.63-1.59 (m, 4H). HRMS (ESI) m/z: calcd for C18H21N2O5S+ [M+H]+, 377.1166; found, 376.8981.
The compound SIAIS171091 was prepared according to the method of intermediate example 26, except that the brominated substrate as the linker was tert-butyl 6-bromohexanoate. The target compound SIAIS171091 was obtained as a white solid (75 mg, total yield of step 3: 53%). 1H NMR (500 MHz, DMSO) δ 11.98 (s, 1H), 10.98 (s, 1H), 7.59-7.52 (m, 3H), 5.12 (dd, J=13.4, 5.1 Hz, 1H), 4.26 (dd, J=40.9, 20.5 Hz, 2H), 3.07 (t, J=7.3 Hz, 2H), 2.96-2.84 (m, 1H), 2.64-2.60 (m, 1H), 2.48-2.39 (m, 1H), 2.19-2.15 (m, 2H), 2.02-2.00 (m, 1H), 1.70-1.35 (m, 6H). HRMS (ESI) m/z: calcd for C19H23N2O5S+ [M+H]+, 391.1322; found, 390.8150.
The compound SIAIS171092 was prepared according to the method of intermediate example 26, except that the brominated substrate as the linker was tert-butyl 7-bromoheptanoate. The target compound SIAIS171092 was obtained as a white solid (79 mg, total yield of step 3: 54%). 1H NMR (500 MHz, DMSO) δ 11.99 (s, 1H), 10.98 (s, 1H), 7.66-7.45 (m, 3H), 5.12 (dd, J=13.3, 5.1 Hz, 1H), 4.26 (dd, J=40.9, 20.5 Hz, 2H), 3.07 (t, J=7.3 Hz, 2H), 2.97-2.83 (m, 1H), 2.63-2.60 (m, 1H), 2.47-2.35 (m, 1H), 2.18 (t, J=7.3 Hz, 2H), 2.06-1.93 (m, 1H), 1.65-1.20 (m, 8H). HRMS (ESI) m/z: calcd for C20H25N2O5S+ [M+H]+, 405.1479; found, 404.8155.
According to Scheme 10, in step 1, the solution of 3-(4-bromo-1-oxoisoindolin-2-yl)piperidine-2,6-dione (0.50 g, 1.5 mmol) in 5 mL DMF was bubbled with argon gas for 5 min. To the mixture were sequentially added but-3-yn-1-ol (0.21 g, 3.0 mmol), Pd(PPh3)2Cl2 (0.10 g, 0.15 mmol) and CuI (57 mg, 0.30 mmol) under stirring for 5 min, followed by addition of 2.5 mL of triethylamine. The mixture was heated to 80° C., and reacted overnight, and then cooled to room temperature. The reaction was quenched with 50 mL of water. The resulting mixture was extracted with ethyl acetate (3×50 mL). The organic phases were combined, washed with water (2×30 mL) and saturated brine (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to remove the solvents. The obtained crude product was subjected to a column chromatography (eluent (v/v): DCM/MeOH=5/1) to give an alcohol intermediate as light yellow solid (0.50 g).
In step 2, to a solution of the above intermediate in 15 mL of DCM were added triethylamine (0.44 g, 4.4 mmol) and Mesyl chloride (0.25 g, 2.2 mmol). The reaction system became clear, and reacted overnight. The reaction mixture was washed with saturated brine, rotary evaporated under reduced pressure to remove the solvents. The resulting residue was subjected to a column chromatography (eluent (v/v): DCM/MeOH=5/1) to give SIAIS255120 as light yellow solid (0.35 g). 1H NMR (500 MHz, DMSO-d6) δ 11.01 (s, 1H), 7.74 (dd, J=7.6, 1.0 Hz, 1H), 7.67 (dd, J=7.6, 1.0 Hz, 1H), 7.54 (t, J=7.6 Hz, 1H), 5.16 (dd, J=13.4, 5.1 Hz, 1H), 4.52-4.27 (m, 4H), 3.24 (s, 3H), 3.02-2.87 (m, 3H), 2.67-2.57 (m, 1H), 2.42 (qd, J=13.3, 4.4 Hz, 1H), 2.03 (m, 1H). HRMS (ESI) m/z: calcd for C18H19N2O6S+ [M+H]+, 391.0958; found, 391.0952.
The compound SIAIS255121 was prepared according to the method of intermediate example 32, except that pent-4-yn-1-ol was used as the starting material. 1H NMR (500 MHz, DMSO-d6) δ 11.00 (s, 1H), 7.72 (dd, J=7.6, 1.0 Hz, 1H), 7.66 (dd, J=7.8, 1.0 Hz, 1H), 7.53 (t, J=7.6 Hz, 1H), 5.15 (dd, J=13.3, 5.1 Hz, 1H), 4.48 (d, J=17.8 Hz, 1H), 4.38-4.28 (m, 3H), 3.20 (s, 3H), 3.00-2.86 (m, 1H), 2.61 m, 3H), 2.45 (dd, J=13.1, 4.5 Hz, 1H), 2.00 (m, 3H). HRMS (ESI) m/z: calcd for C19H21N2O6S+ [M+H]+, 405.1115; found, 405.1111.
The compound SIAIS255119 was prepared according to the method of intermediate example 32, except that hex-5-yn-1-ol was used as the starting material. 1H NMR (500 MHz, DMSO-d6) δ 10.99 (s, 1H), 7.71 (dd, J=7.6, 1.1 Hz, 1H), 7.65 (dd, J=7.7, 1.0 Hz, 1H), 7.52 (t, J=7.6 Hz, 1H), 5.14 (dd, J=13.4, 5.1 Hz, 1H), 4.46 (d, J=17.7 Hz, 1H), 4.31 (d, J=17.7 Hz, 1H), 4.27 (t, J=6.4 Hz, 2H), 3.17 (s, 3H), 2.91 (m, 1H), 2.55 (m, 3H), 2.48-2.42 (m, 1H), 2.01 (m, 1H), 1.88-1.80 (m, 2H), 1.67 (m, 2H). HRMS (ESI) m/z: calcd for C20H23N2O6S+ [M+H]+, 419.1271; found, 419.1270.
According to Scheme 11, in step 1, a 10 mL reaction flask was sequentially charged with the compound SIAIS171095 (0.36 mmol, 1 equiv), anhydrous DMF (2 mL) and anhydrous potassium carbonate (0.72 mmol, 2 equiv) with stirring at room temperature, followed by slow addition of the corresponding tert-butyl (2-bromoethyl)carbamate (0.43 mmol, 1.2 equiv). After the completion of addition, the mixture was stirred at room temperature for 1 h. After the starting materials were consumed, the crude product was separated by a reverse phase C18 column chromatography (eluent (v/v): acetonitrile/(water+0.05% TFA)=10%-100%), and the collected fractions were rotary evaporated under reduced pressure to remove the solvents. The resulting residue was lyophilized to give the Boc-protected alkylated intermediate product.
The corresponding intermediate compound obtained from step 1, dichloromethane (2 mL) and trifluoroacetic acid (2 mL) were sequentially added to a 10 mL reaction flask, and stirred at room temperature for 12 h, and then rotary evaporated under reduced pressure to remove the solvents. The resulting crude product was separated by a reverse phase C18 column chromatography (eluent (v/v): acetonitrile/(water+0.05% TFA)=10%-100%), and the collected fractions were rotary evaporated under reduced pressure to remove the solvents. The resulting residue was lyophilized to give the target product SIAIS171123 as white solid (68 mg, total yield of two steps 59%). 1H NMR (500 MHz, DMSO) δ 11.02 (s, 1H), 7.88 (s, 3H), 7.73 (dd, J=7.7, 0.8 Hz, 1H), 7.66 (dd, J=7.5, 0.7 Hz, 1H), 7.59 (t, J=7.6 Hz, 1H), 5.15 (dd, J=13.3, 5.1 Hz, 1H), 4.45-4.25 (m, 2H), 3.32-3.26 (m, 2H), 3.05-3.00 (m, 2H), 2.96-2.87 (m, 1H), 2.64-2.60 (m, 1H), 2.48-2.41 (m, 1H), 2.05-2.00 (m, 1H). HRMS (ESI) m/z: calcd for C15H18N3O3S+ [M+H]+, 320.1063; found, 320.1082.
The compound SIAIS171124 was prepared according to the method of intermediate example 35, except that the brominated substrate as the linker was tert-butyl (3-bromobutyl)carbamate. The target compound SIAIS171124 was obtained as a white solid (68 mg, total yield of two steps 56%). 1H NMR (500 MHz, DMSO) δ 11.00 (s, 1H), 7.75-7.67 (m, 4H), 7.63-7.49 (m, 2H), 5.14 (dd, J=13.3, 5.1 Hz, 1H), 4.43-4.16 (m, 2H), 3.22-3.11 (m, 2H), 2.97-2.85 (m, 3H), 2.67-2.56 (m, 1H), 2.48-2.40 (m, 1H), 2.05-1.95 (m, 1H), 1.91-1.77 (m, 2H). HRMS (ESI) m/z: calcd for C16H20N3O3S+ [M+H]+, 334.1220; found, 334.1213.
The compound SIAIS171131 was prepared according to the method of intermediate example 35, except that the brominated substrate as the linker was tert-butyl (4-bromobutyl)carbamate. The target compound SIAIS171131 was obtained as a light yellow solid (76 mg, total yield of two steps 60%). 1H NMR (500 MHz, DMSO) δ 10.99 (s, 1H), 7.81-7.47 (m, 6H), 5.13 (dd, J=13.3, 5.1 Hz, 1H), 4.25 (dd, J=31.3, 15.7 Hz, 2H), 3.20-3.03 (m, 2H), 2.96-2.85 (m, 1H), 2.85-2.80 (m, 2H), 2.63-2.60 (m, 1H), 2.46-2.30 (m, 1H), 2.06-1.94 (m, 1H), 1.71-1.56 (m, 4H). HRMS (ESI) m/z: calcd for C17H22N3O3S+ [M+H]+, 348.1376; found, 348.1381.
The compound SIAIS171132 was prepared according to the method of intermediate example 35, except that the brominated substrate as the linker was tert-butyl (5-bromopentyl)carbamate. The target compound SIAIS171132 was obtained as a light yellow solid (95 mg, total yield of two steps 73%). 1H NMR (500 MHz, DMSO) δ 11.00 (s, 1H), 7.85-7.45 (m, 6H), 5.21-5.07 (m, 1H), 4.42-4.16 (m, 2H), 3.16-3.05 (m, 2H), 2.92-2.85 (m, 1H), 2.84-2.71 (m, 2H), 2.64-2.60 (m, 1H), 2.45-2.40 (m, 1H), 2.07-1.93 (m, 1H), 1.66-1.58 (m, 2H), 1.54-1.50 (m, 2H), 1.49-1.44 (m, 2H). HRMS (ESI) m/z: calcd for C18H24N3O3S+ [M+H]+, 362.1533; found, 362.1537.
The compound SIAIS171134 was prepared according to the method of intermediate example 35, except that the brominated substrate as the linker was tert-butyl (6-bromohexyl)carbamate. The target compound SIAIS171134 was obtained as a light yellow solid (78 mg, total yield of two steps 57%). 1H NMR (500 MHz, DMSO) δ 11.00 (s, 1H), 7.84-7.43 (m, 6H), 5.16-5.13 (m, 1H), 4.30-4.15 (m, 2H), 3.14-3.03 (m, 2H), 2.97-2.88 (m, 1H), 2.82-2.72 (m, 2H), 2.62 (t, J=14.7 Hz, 1H), 2.49-2.39 (m, 1H), 2.06-1.96 (m, 1H), 1.68-1.56 (m, 2H), 1.51-1.46 (m, 2H), 1.45-1.37 (m, 2H), 1.36-1.28 (m, 2H). HRMS (ESI) m/z: calcd for C19H26N3O3S+ [M+H]+, 376.1689; found, 376.1702.
The compound SIAIS171135 was prepared according to the method of intermediate example 35, except that the brominated substrate as the linker was tert-butyl (7-bromoheptyl)carbamate. The target compound SIAIS171135 was obtained as a white solid (100 mg, total yield of two steps 71%). 1H NMR (500 MHz, DMSO) δ 10.99 (s, 1H), 7.84-7.42 (m, 6H), 5.13 (dd, J=13.3, 5.1 Hz, 1H), 4.37-4.18 (m, 2H), 3.15-3.02 (m, 2H), 2.92-2.88 (m, 1H), 2.81-2.71 (m, 2H), 2.61 (t, J=14.8 Hz, 1H), 2.48-2.40 (m, 1H), 2.05-1.98 (m, 1H), 1.65-1.56 (m, 2H), 1.54-1.46 (m, 2H), 1.44-1.36 (m, 2H), 1.33-1.23 (m, 4H). HRMS (ESI) m/z: calcd for C20H28N3O3S+ [M+H]+, 390.1846; found, 390.1846.
The compound SIAIS171136 was prepared according to the method of intermediate example 35, except that the brominated substrate as the linker was tert-butyl (8-bromooctyl)carbamate. The target compound SIAIS171136 was obtained as a white solid (100 mg, total yield of two steps 68%). 1H NMR (500 MHz, DMSO) δ 10.99 (s, 1H), 7.75-7.47 (m, 6H), 5.13 (dd, J=13.3, 5.1 Hz, 1H), 4.28 (dd, J=70.1, 17.4 Hz, 2H), 3.13-3.00 (m, 2H), 2.98-2.84 (m, 1H), 2.78-2.74 (m, 2H), 2.64-2.59 (m, 1H), 2.47-2.38 (m, 1H), 2.06-1.93 (m, 1H), 1.68-1.54 (m, 2H), 1.52-1.48 (m, 2H), 1.45-1.34 (m, 2H), 1.30-1.20 (m, 6H). HRMS (ESI) m/z: calcd for C21H30N3O3S+ [M+H]+, 404.2002; found, 404.1996.
Referring to Scheme 12, a 50 mL two-necked flask was sequentially charged with the compound SIAIS171095 (0.344 mmol, 1 equiv), potassium carbonate (0.688 mmol, 2 equiv), and DMF (5 mL), followed by evacuation and refilling with argon gas, and addition of 1,6-dibromohexane (0.413 mmol, 1.2 equiv). The mixture was stirred and reacted at room temperature for 1 h. After the reaction was complete, the mixture was filtered to remove the insoluble substance, and the filtrate was then subjected to a reverse phase C18 column chromatography (eluent (v/v): acetonitrile/(water)=10%-100%) for separation, and the collected fractions were concentrated under reduced pressure to remove the solvents, to give the corresponding target compound SIAIS1216133 (white solid, 339 mg, yield 38%). 1H NMR (500 MHz, DMSO) δ 11.01 (s, 1H), 7.63 (dd, J=7.5, 1.2 Hz, 1H), 7.58-7.51 (m, 2H), 5.13 (dd, J=13.3, 5.1 Hz, 1H), 4.35 (d, J=17.4 Hz, 1H), 4.21 (d, J=17.4 Hz, 1H), 3.52 (t, J=6.7 Hz, 2H), 3.08 (t, J=7.2 Hz, 2H), 2.96-2.87 (m, 1H), 2.59 (d, J=17.4 Hz, 1H), 2.49-2.41 (m, 1H), 2.04-1.97 (m, 1H), 1.82-1.74 (m, 2H), 1.63-1.56 (m, 2H), 1.46-1.36 (m, 4H). HRMS (ESI) m/z: calcd for C19H24BrN2O3S+ [M+H]+, 439.0686; found, 439.0680.
According to Scheme 13, in step 1, to the solution of 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindolin-1,3-dione (16.8 mmol, 1 equiv) dissolve in 25 mL NMP were sequentially added 6-aminohexan-1-ol (16.8 mmol, 1.0 equiv) and DMF (25.2 mmol, 1.5 equiv). The mixture was heated and reacted at 90° C. for 4 h. After the reaction was complete, the reaction mixture was cooled to room temperature, and poured into saturated brine. The resulting mixture was extracted with ethyl acetate (4×50 mL). The organic phases were combined, washed with water (2×30 mL) and saturated brine (50 mL), dried over anhydrous sodium sulfate, and rotary evaporated under reduced pressure to remove the solvents. The obtained crude product was subjected to a column chromatography (eluent (v/v): petroleum ether/ethyl acetate=1:1) for purification, to give the intermediate. To a solution of the intermediate dissolved in 50 mL of tetrahydrofuran was added tetrabutylammonium fluoride (16.8 mmol). The resulting mixture was stirred at room temperature for 2 h. After the reaction was complete, to the mixture was added saturated brine (200 mL), followed by extraction with ethyl acetate (4×50 mL). The organic phases were combined, washed with water (2×30 mL) and saturated brine (50 mL), dried over anhydrous sodium sulfate, and rotary evaporated under reduced pressure to remove the solvents, to give the crude product (1.0 g) which was used directly in the next step.
In step 2, to the solution of the crude product from step 1 dissolved in 40 mL of mixed solvent (DCM/pyridine=3/1) were sequentially added triethylamine (0.52 mL, 3.8 mmol) and methanesulfonyl chloride (0.30 mL, 3.8 mmol). The mixture was heated to 40° C., and reacted for 2 h. After the reaction was complete, the mixture was washed with saturated brine, and rotary evaporated under reduced pressure to remove the solvents. The resulting residue was subjected to a column chromatography to give a yellow powder (m=0.80 g).
In step 3, to the solution of the crude product from step 2 dissolved in 10 mL of acetone was added sodium iodide (3.0 equiv). The mixture was heated to 60° C., and reacted for 24 h. After the conversion was complete, the mixture was cooled to room temperature, and diluted with 40 mL ethyl acetate. The resulting mixture was washed with saturated brine, dried over anhydrous sodium sulfate, and rotary evaporated under reduced pressure to remove the solvents, to give the product SIAIS264018 as a yellow solid which was used directly in the next step. 1H NMR (500 MHz, DMSO-d6) δ 11.09 (s, 1H), 7.58 (dd, J=8.5, 7.0 Hz, 1H), 7.10 (d, J=8.6 Hz, 1H), 7.02 (d, J=7.0 Hz, 1H), 6.54 (t, J=5.9 Hz, 1H), 5.05 (dd, J=12.7, 5.4 Hz, 1H), 3.28 (q, J=6.7 Hz, 4H), 2.95-2.83 (m, 1H), 2.63-2.55 (m, 1H), 2.08 (d, J=4.9 Hz, 1H), 2.06-1.99 (m, 1H), 1.77 (t, J=7.0 Hz, 2H), 1.57 (t, J=7.1 Hz, 2H), 1.38 (p, J=5.0 Hz, 4H).
Osimertinib derivative (SIAIS337051) was prepared according to Scheme 16.
Steps 1 and 2:
An egg-shaped flask was sequentially charged with N-(4-fluoro-2-methoxy-5-nitrophenyl)-4-(1-methyl-1H-indol-3-yl)pyrimidin-2-amine (1.18 g, 3 mmol), DMF (6 mL), tert-butyl methyl(2-(methylamino)ethyl)carbamate (677.8 mg, 3.6 mmol), and potassium carbonate (621.9 mg, 4.5 mmol) at room temperature, followed by evacuation and refilling with argon gas and reacting at 80° C. for 12 h. After the reaction was complete as monitored by TLC, the reaction was quenched with water. The mixture was extracted with ethyl acetate, and rotary evaporated. The resulting residue was subjected to a column chromatography (eluent (v/v): DCM:MeOH=30:1) for separation, to give a yellow solid (900 mg) which was used directly in the next step. To a solution of the obtained yellow solid dissolved in 20 mL of 75% ethanol were added iron powder (268.8 mg, 4.8 mmol) and ammonium chloride (342 mg, 6.4 mmol), followed by evacuation and refilling with argon gas and reacting at 80° C. for 3 h. After the reaction was complete as monitored by TLC, the mixture was filtered by a Buchner funnel. The filtrate cake was washed with a mixed solvent (dichloromethane:methanol=10:1). The filtrate was rotary evaporated, and the resulting residue was subjected to a column chromatography (eluent (v/v): DCM:MeOH=30:1) for separation, to give a yellow solid (780 mg, total yield of two steps 49%). HRMS (ESI) m/z: calcd for C29H36N7O5+ [M+H]+, 562.2772; found, 562.2771.
Steps 3 and 4:
A 100 mL clean egg-shaped flask was sequentially charged with the yellow solid obtained from the previous step (300 mg, 0.565 mmol), and DMF (5 mL), followed by addition of 3-chloropropanoyl chloride (60 μL, 0.633 mmol) with stirring at 0° C. The mixture was warmed up to room temperature, stirred and reacted at room temperature for 1 h. After the reaction was complete, the reaction was quenched with water. The resulting mixture was extracted with dichloromethane, washed with saturated brine, dried over anhydrous sodium sulfate, and rotary evaporated. The resulting residue was used directly in the next step. To a solution of the product obtained from the previous step (0.565 mmol) in 5 mL of acetonitrile was added triethylamine (171.2 mg, 1.695 mmol). The resulting mixture was reacted at 80° C. for 12 h. After the reaction was complete as monitored by LC-MS, the mixture was rotary evaporated, and the resulting residue was subjected to a silica gel column chromatography (eluent (v/v): dichloromethane/methanol=30:1) for separation, to give a yellow solid (280 mg, total yield of two steps 82%). HRMS (ESI) m/z: calcd for C32H40N7O4+ [M+H]+, 586.3136; found, 586.3133.
Step 5:
To a solution of the yellow solid obtained from the previous step dissolved in 2 mL of MeOH was added 4 mL of a solution of hydrochloric acid in dioxane (4M). The mixture was reacted at room temperature for 2 h. After the reaction was complete as monitored by LC-MS, the mixture was rotary evaporated, and the resulting residue was subjected to C18 reverse phase column chromatography (eluent: MeOH/water) for separation to give the SIAIS337051 as a yellow solid (228 mg, yield 99%). 1H NMR (500 MHz, DMSO-d6) δ 9.83 (s, 1H), 9.13 (s, 2H), 8.80 (s, 1H), 8.32 (d, J=89.5 Hz, 2H), 7.59 (d, J=8.2 Hz, 1H), 7.40 (d, J=6.6 Hz, 1H), 7.27 (dt, J=31.2, 7.7 Hz, 2H), 7.13 (dd, J=16.9, 10.2 Hz, 1H), 7.00 (s, 1H), 6.21 (d, J=16.9 Hz, 1H), 5.71 (d, J=10.3 Hz, 1H), 3.93 (s, 3H), 3.84 (s, 3H), 3.32 (d, J=5.6 Hz, 2H), 3.15 (q, J=6.0, 5.6 Hz, 2H), 2.64 (s, 3H), 2.58 (t, J=5.5 Hz, 3H). HRMS (ESI) m/z: calcd for C27H32N7O2+ [M+H]+, 486.2612; found, 486.2614.
Referring to Scheme 1, Dacomitinib derivative B was prepared by using a method similar to that of Dacomitinib derivative A in Intermediate Example 1. The synthetic and structural characterization data of the intermediate are as follows:
(E)-4-(4-aminopiperidin-1-yl)-N-(4-((3-chloro-4-fluorophenyl)amino)-7-methoxyquinazolin-6-yl)but-2-enamide (SIAIS249077). (yellow solid, 310 mg, total yield of two steps 73%) 1H NMR (500 MHz, MeOD) δ 9.19 (s, 1H), 8.70 (s, 1H), 7.94 (dd, J=6.7, 2.6 Hz, 1H), 7.69-7.63 (m, 1H), 7.36 (d, J=8.9 Hz, 1H), 7.32 (d, J=13.5 Hz, 1H), 7.06 (dt, J=14.7, 7.0 Hz, 1H), 6.81 (d, J=15.2 Hz, 1H), 4.16 (s, 3H), 3.92 (s, 1H), 3.59 (s, 1H), 3.08-2.99 (m, 1H), 2.26 (d, J=13.7 Hz, 2H), 2.19 (t, J=7.5 Hz, 1H), 2.09-1.98 (m, 2H), 1.65-1.58 (m, 1H), 1.32 (s, 2H). HRMS (ESI) m/z: calcd for C24H27ClFN6O2+ [M+H]+:485.1863, found 485.1869.
The compound SIAIS255127 was prepared according to the method of intermediate example 32, except that the non-8-yn-1-ol was used as the starting material. 1H NMR (500 MHz, DMSO) δ 11.00 (s, 1H), 7.70 (d, J=7.4 Hz, 1H), 7.63 (d, J=7.0 Hz, 1H), 7.52 (t, J=7.6 Hz, 1H), 5.14 (dd, J=13.3, 5.1 Hz, 1H), 4.44 (d, J=17.6 Hz, 1H), 4.30 (d, J=17.6 Hz, 1H), 4.18 (t, J=6.5 Hz, 2H), 3.15 (s, 3H), 2.91 (ddd, J=17.5, 13.7, 5.4 Hz, 1H), 2.63-2.57 (m, 1H), 2.47 (d, J=7.1 Hz, 2H), 2.46-2.40 (m, 1H), 2.02 (ddd, J=10.3, 5.1, 3.1 Hz, 1H), 1.66 (dd, J=13.5, 6.6 Hz, 2H), 1.61-1.53 (m, 2H), 1.47-1.40 (m, 2H), 1.40-1.33 (m, 4H). HRMS (ESI) m/z: calcd for C23H29N2O6S+ [M+H]+, 461.1741; found, 461.1740.
A 50 mL egg-shaped flask was sequentially charged with Lenalidomide (1 mmol, 1 equiv), anhydrous N,N-dimethylformamide (10 mL) and DIPEA (3 mmol, 3 equiv), followed by addition of 1,4-bis(bromomethyl)benzene (2 mmol, 2 equiv) with stirring at room temperature. The mixture was stirred at 40° C. for 3 h. After the reaction was complete, the reaction mixture was filtered to remove the insoluble substance, and the filtrate was then directly subjected to a reverse phase C18 column chromatography (eluent (v/v): acetonitrile/water=10%-100%) for separation, and the collected fractions were concentrated under reduced pressure to remove the solvents, to give the corresponding product SIAIS1221131 as a yellow solid (235 mg, yield 53%). 1H NMR (500 MHz, DMSO) δ 11.02 (s, 1H), 7.38 (q, J=8.3 Hz, 4H), 7.20 (t, J=7.7 Hz, 2H), 6.93 (d, J=7.3 Hz, 1H), 6.62 (d, J=8.1 Hz, 1H), 5.12 (dd, J=13.3, 5.0 Hz, 1H), 4.68 (s, 2H), 4.39 (s, 2H), 4.32 (d, J=17.2 Hz, 1H), 4.20 (d, J=17.1 Hz, 1H), 2.97-2.89 (m, 1H), 2.63 (d, J=16.1 Hz, 1H), 2.38-2.27 (m, 1H), 2.09-2.01 (m, 1H). HRMS (ESI) m/z: calcd for C21H21BrN3O3+ [M+H]+, 442.0761; found, 442.0766.
A 50 mL egg-shaped flask was sequentially charged with hydroxy-substituted lenalidomide (CAS No.: 1061604-41-8; 2 mmol, 1 equiv), acetonitrile (10 mL) and potassium carbonate (4 mmol, 2 equiv), followed by addition of tert-butyl 2-bromoacetate (2.4 mmol, 1.2 equiv) with stirring at room temperature. The mixture was reacted at 80° C. for 4 h. After the reaction was complete, the reaction mixture was concentrated under reduced pressure to remove acetonitrile. The resulting residue was dissolved in a small amount of DMSO, and subjected to a reverse phase C18 column chromatography (eluent (v/v): acetonitrile/water=10%-100%) for separation, and the collected fractions were concentrated under reduced pressure to remove the solvents, to give the corresponding tert-butyl ester intermediate compound. The corresponding tert-butyl ester intermediate compound, dichloromethane (1 mL) and trifluoroacetic acid (3 mL) were sequentially added to a 25 mL egg-shaped flask, and stirred at room temperature for 1 h, and then concentrated under reduced pressure to remove the solvents. The resulting residue was treated by addition of water and lyophilized to afford the corresponding target compound SIAIS1222121 (light yellow solid, 371 mg, yield 58%). 1H NMR (500 MHz, DMSO) δ 10.99 (s, 1H), 7.48 (t, J=7.6 Hz, 1H), 7.35 (d, J=7.6 Hz, 1H), 7.17 (d, J=8.2 Hz, 1H), 5.12 (dd, J=13.5, 4.9 Hz, 1H), 4.86 (s, 2H), 4.41 (d, J=17.5 Hz, 1H), 4.27 (d, J=17.4 Hz, 1H), 2.96-2.88 (m, 1H), 2.60 (d, J=17.1 Hz, 1H), 2.48-2.42 (m, 1H), 2.04-1.97 (m, 1H). HRMS (ESI) m/z: calcd for C15H15N2O6+ [M+H]+, 319.0925; found, 319.0928.
The compound SIAIS122212 was prepared according to the method of Intermediate Example 48, except that the brominated substrate as the linker was tert-butyl 5-bromopentanoate. The target compound SIAIS122212 was obtained as a light yellow solid (418 mg, yield: 59%). 1H NMR (500 MHz, DMSO) δ 10.97 (s, 1H), 7.48 (t, J=7.6 Hz, 1H), 7.31 (d, J=7.5 Hz, 1H), 7.24 (d, J=8.0 Hz, 1H), 5.11 (dd, J=13.3, 5.0 Hz, 1H), 4.37 (d, J=17.3 Hz, 1H), 4.23 (d, J=17.3 Hz, 1H), 4.13 (t, J=5.9 Hz, 2H), 2.96-2.87 (m, 1H), 2.59 (d, J=17.2 Hz, 1H), 2.49-2.42 (m, 1H), 2.30 (t, J=7.1 Hz, 2H), 2.02-1.95 (m, 1H), 1.79-1.75 (m, 2H), 1.73-1.64 (m, 2H). HRMS (ESI) m/z: calcd for C18H21N2O6+ [M+H]+, 361.1394; found, 361.1391.
The compound SIAIS1222149 was prepared according to the method of Intermediate Example 48, except that the brominated substrate as the linker was tert-butyl 6-bromohexanoate. The target compound SIAIS1222149 was obtained as a light yellow solid (313 mg, yield: 42%). 1H NMR (500 MHz, DMSO) δ 10.97 (s, 1H), 7.48 (t, J=7.7 Hz, 1H), 7.31 (d, J=7.5 Hz, 1H), 7.24 (d, J=8.1 Hz, 1H), 5.11 (dd, J=13.3, 4.9 Hz, 1H), 4.38 (d, J=17.4 Hz, 1H), 4.23 (d, J=17.3 Hz, 1H), 4.11 (t, J=6.2 Hz, 2H), 2.97-2.87 (m, 1H), 2.59 (d, J=17.6 Hz, 1H), 2.49-2.40 (m, 1H), 2.24 (t, J=7.3 Hz, 2H), 2.03-1.96 (m, 1H), 1.79-1.72 (m, 2H), 1.61-1.54 (m, 2H), 1.50-1.41 (m, 2H). HRMS (ESI) m/z: calcd for C19H23N2O6+ [M+H]+, 375.1551; found, 375.1535.
The compound SIAIS1222151 was prepared according to the method of Intermediate Example 48, except that the brominated substrate as the linker was tert-butyl 7-bromoheptanoate. The target compound SIAIS1222151 was obtained as a light yellow solid (250 mg, yield: 32%). 1H NMR (500 MHz, DMSO) δ 10.97 (s, 1H), 7.48 (t, J=7.8 Hz, 1H), 7.31 (d, J=7.4 Hz, 1H), 7.24 (d, J=8.1 Hz, 1H), 5.11 (dd, J=13.2, 4.9 Hz, 1H), 4.38 (d, J=17.3 Hz, 1H), 4.23 (d, J=17.3 Hz, 1H), 4.11 (t, J=6.2 Hz, 2H), 2.95-2.87 (m, 1H), 2.59 (d, J=18.3 Hz, 1H), 2.49-2.41 (m, 1H), 2.22 (t, J=7.3 Hz, 2H), 2.03-1.96 (m, 1H), 1.77-1.70 (m, 2H), 1.56-1.49 (m, 2H), 1.48-1.40 (m, 2H), 1.38-1.30 (m, 2H). HRMS (ESI) m/z: calcd for C20H25N2O6+[M+H]+, 389.1707; found, 389.1702.
A 50 mL egg-shaped flask was sequentially charged with hydroxy-substituted lenalidomide (CAS No.: 1061604-41-8; 2 mmol, 1 equiv), acetonitrile (10 mL) and potassium carbonate (4 mmol, 2 equiv), followed by addition of 1,6-dibromohexane (4 mmol, 2 equiv) with stirring at room temperature. The mixture was reacted at 80° C. overnight. After the reaction was complete, the reaction mixture was concentrated under reduced pressure to remove acetonitrile. The resulting residue was dissolved in a small amount of DMSO, and subjected to a reverse phase C18 column chromatography (eluent (v/v): acetonitrile/water=10%-100%) for separation, and the collected fractions were concentrated under reduced pressure to remove the solvents, to give the corresponding target compound SIAIS1222127 (light yellow solid, 318 mg, yield 38%). 1H NMR (500 MHz, DMSO) δ 10.91 (s, 1H), 7.48 (t, J=7.8 Hz, 1H), 7.31 (d, J=7.4 Hz, 1H), 7.24 (d, J=8.1 Hz, 1H), 5.11 (dd, J=13.3, 5.0 Hz, 1H), 4.38 (d, J=17.4 Hz, 1H), 4.23 (d, J=17.3 Hz, 1H), 4.12 (t, J=6.2 Hz, 2H), 3.54 (t, J=6.6 Hz, 2H), 2.96-2.86 (m, 1H), 2.59 (d, J=17.4 Hz, 1H), 2.47-2.40 (m, 1H), 2.03-1.96 (m, 1H), 1.88-1.79 (m, 2H), 1.78-1.72 (m, 2H), 1.46 (s, 4H). HRMS (ESI) m/z: calcd for C19H24BrN2O4+ [M+H]+, 423.0914; found, 423.0913.
Referring to Scheme 14, a reaction flask was sequentially charged with the corresponding EGFR inhibitor, i.e., Dacomitinib derivative A (0.02 mmol, 1 equiv), intermediate LM (SIAIS151001) (0.02 mmol, 1 equiv), HOAt (0.04 mmol, 2 equiv), EDCI (0.04 mmol, 2 equiv), 2 mL DMF, and NMM (0.2 mmol, 10 equiv) at room temperature, and the reaction mixture was reacted overnight. After the reaction was complete as monitored by TLC, the reaction mixture was filtered, and the filtrate was subjected to preparative HPLC (eluent (v/v): acetonitrile/(water+0.05% HCl)=10%-100%) for separation, and the collected fractions were rotary evaporated under reduced pressure to remove the acetonitrile. The resulting residue was lyophilized to give the final target compound (SIAIS249046). (yellow solid, 8.9 mg, yield 50%)1H NMR (500 MHz, MeOD) δ 9.23 (s, 1H), 8.76 (s, 1H), 7.95 (dd, J=6.6, 2.5 Hz, 1H), 7.70-7.65 (m, 1H), 7.47 (s, 1H), 7.39 (t, J=8.9 Hz, 1H), 7.30 (s, 1H), 7.08-6.99 (m, 2H), 6.94 (d, J=6.9 Hz, 1H), 6.79 (d, J=15.2 Hz, 1H), 5.09 (dd, J=12.7, 5.5 Hz, 1H), 4.16 (s, 3H), 4.04 (d, J=6.5 Hz, 2H), 3.81 (s, 2H), 3.74 (d, J=5.1 Hz, 1H), 3.72 (d, J=5.2 Hz, 1H), 3.71-3.32 (m, 8H), 2.94-2.84 (m, 1H), 2.75-2.68 (m, 2H), 2.17-2.13 (m, 1H). HRMS (ESI) m/z: calcd for C41H42ClFN9O8+[M+H]+, 842.2823; found, 842.2820.
Referring to the method of example 1, the target compound (SIAIS262013) was prepared by using Dacomitinib derivative A and intermediate LM (SIAIS151004). (yellow solid, 8.2 mg, yield 44%)1H NMR (500 MHz, MeOD) δ 9.24 (s, 1H), 8.78 (s, 1H), 7.96 (dd, J=6.6, 2.5 Hz, 1H), 7.72-7.65 (m, 1H), 7.49 (s, 1H), 7.41 (t, J=8.9 Hz, 1H), 7.32 (s, 1H), 7.09-6.99 (m, 2H), 6.95 (d, J=6.9 Hz, 1H), 6.81 (d, J=15.2 Hz, 1H), 5.09 (dd, J=12.7, 5.5 Hz, 1H), 4.18 (s, 3H), 4.03 (d, J=6.5 Hz, 2H), 3.84 (s, 2H), 3.74 (d, J=5.1 Hz, 1H), 3.71 (d, J=5.2 Hz, 1H), 3.72-3.34 (m, 8H), 3.31-3.23 (m, 4H), 2.96-2.85 (m, 1H), 2.75-2.68 (m, 2H), 2.17-2.13 (m, 1H). HRMS (ESI) m/z: calcd for C43H46ClFN9O9+ [M+H]+, 886.3086; found, 886.3083.
Referring to the method of example 1, the target compound (SIAIS249047) was prepared by using Dacomitinib derivative A and intermediate LM (SIAIS151005). (yellow solid, 9.9 mg, yield 51%)1H NMR (500 MHz, MeOD) δ 9.22 (s, 1H), 8.75 (s, 1H), 7.94 (dd, J=6.6, 2.5 Hz, 1H), 7.71-7.62 (m, 1H), 7.50-7.42 (m, 1H), 7.37 (t, J=8.9 Hz, 1H), 7.32 (s, 1H), 7.09-7.01 (m, 2H), 6.94 (d, J=7.0 Hz, 1H), 6.83 (d, J=15.2 Hz, 1H), 5.06 (dd, J=12.8, 5.5 Hz, 1H), 4.17 (s, 3H), 4.08 (d, J=7.1 Hz, 2H), 3.78-3.71 (m, 4H), 3.69-3.65 (m, 6H), 3.60 (d, J=3.3 Hz, 4H), 3.48 (t, J=5.0 Hz, 2H), 3.22-3.00 (m, 2H), 2.91-2.84 (m, 1H), 2.73-2.68 (m, 2H), 2.17-2.08 (m, 1H). HRMS (ESI) m/z: calcd for C45H50ClFN9O10+ [M+H]+, 930.3348; found, 930.3344.
Referring to the method of example 1, the target compound (SIAIS262014) was prepared by using Dacomitinib derivative A and intermediate LM (SIAIS151006). (yellow solid, 10.1 mg, yield 49%)1H NMR (500 MHz, MeOD) δ 9.19 (s, 1H), 8.73 (s, 1H), 7.93 (dd, J=6.6, 2.6 Hz, 1H), 7.68-7.64 (m, 1H), 7.43 (dd, J=8.5, 7.2 Hz, 1H), 7.37 (t, J=8.9 Hz, 1H), 7.30 (s, 1H), 7.04 (dd, J=15.1, 7.4 Hz, 1H), 6.98 (d, J=8.6 Hz, 1H), 6.90 (d, J=7.0 Hz, 1H), 6.85 (d, J=15.2 Hz, 1H), 5.07 (dd, J=12.6, 5.5 Hz, 1H), 4.16 (s, 3H), 4.12-4.04 (m, 2H), 3.73 (t, J=5.8 Hz, 6H), 3.67 (s, 2H), 3.65-3.57 (m, 8H), 3.46-3.43 (m, 2H), 2.91-2.86 (m, 2H), 2.79-2.72 (m, 2H), 2.72-2.63 (m, 1H), 2.14-2.10 (m, 1H). HRMS (ESI) m/z: calcd for C47H54ClFN9O11+ [M+H]+, 974.3610; found, 974.3612.
Referring to the method of example 1, the target compound (SIAIS219194) was prepared by using Dacomitinib derivative A and intermediate LM (SIAIS151025). (yellow solid, 8.2 mg, yield 50%)1H NMR (500 MHz, DMSO) δ 11.10 (s, 1H), 9.81 (s, 1H), 9.72 (s, 1H), 8.92 (s, 1H), 8.53 (s, 1H), 8.15-8.09 (m, 1H), 7.83-7.76 (m, 1H), 7.64-7.58 (m, 1H), 7.42 (t, J=9.1 Hz, 1H), 7.29 (s, 1H), 7.09 (d, J=4.2 Hz, 2H), 6.85-6.80 (m, 1H), 6.62 (d, J=15.3 Hz, 1H), 5.06 (dd, J=12.8, 5.4 Hz, 1H), 4.20 (d, J=4.1 Hz, 2H), 4.02 (s, 3H), 3.55 (d, J=16.3 Hz, 8H), 3.21 (d, J=5.8 Hz, 2H), 2.65-2.57 (m, 1H), 2.42 (s, 2H), 2.07-2.00 (m, 1H). HRMS (ESI) m/z: calcd for C38H35ClFN9O7+ [M+H]+, 783.2332; found, 783.2330.
Referring to the method of example 1, the target compound (SIAIS262016) was prepared by using Dacomitinib derivative A and intermediate LM (SIAIS151026). (yellow solid, 6.6 mg, yield 39%)1H NMR (500 MHz, MeOD) δ 9.25 (s, 1H), 8.74 (s, 1H), 7.93 (dd, J=6.6, 2.6 Hz, 1H), 7.68-7.63 (m, 1H), 7.58 (dd, J=8.5, 7.2 Hz, 1H), 7.37 (t, J=8.9 Hz, 1H), 7.32 (s, 1H), 7.13 (d, J=8.6 Hz, 1H), 7.08-7.00 (m, 2H), 6.83 (d, J=15.3 Hz, 1H), 5.07 (dd, J=12.6, 5.5 Hz, 1H), 4.17 (d, J=5.4 Hz, 3H), 4.01 (d, J=7.1 Hz, 2H), 3.74-3.66 (m, 2H), 3.30 (s, 8H), 2.92-2.69 (m, 5H), 2.16-2.08 (m, 1H). HRMS (ESI) m/z: calcd for C39H37ClFN9O7+ [M+H]+, 797.2489; found, 797.2485.
Referring to the method of example 1, the target compound (SIAIS249062) was prepared by using Dacomitinib derivative A and intermediate LM (SIAIS151019). (yellow solid, 8.2 mg, yield 48%)1H NMR (500 MHz, MeOD) δ 9.29 (s, 1H), 8.76 (s, 1H), 7.93 (dd, J=6.6, 2.5 Hz, 1H), 7.66 (dd, J=4.8, 2.7 Hz, 1H), 7.57 (dd, J=8.5, 7.2 Hz, 1H), 7.38 (t, J=8.9 Hz, 1H), 7.34 (s, 1H), 7.13 (d, J=8.5 Hz, 1H), 7.07 (dt, J=11.1, 5.7 Hz, 2H), 6.87 (d, J=15.2 Hz, 1H), 5.07 (dd, J=12.6, 5.4 Hz, 1H), 4.19 (s, 3H), 4.07 (d, J=7.1 Hz, 2H), 3.43 (t, J=6.2 Hz, 2H), 3.32-3.30 (m, 8H), 2.90-2.80 (m, 1H), 2.77-2.72 (m, 2H), 2.57 (s, 2H), 2.12 (d, J=5.2 Hz, 1H), 2.01 (d, J=10.9 Hz, 2H). HRMS (ESI) m/z: calcd for C40H39ClFN9O7+ [M+H]+, 811.2645; found, 811.2642.
Referring to the method of example 1, the target compound (SIAIS249048) was prepared by using Dacomitinib derivative A and intermediate LM (SIAIS151020). (yellow solid, 8.1 mg, yield 47%)1H NMR (500 MHz, MeOD) δ 9.22 (s, 1H), 8.70 (s, 1H), 7.88 (dd, J=6.6, 2.5 Hz, 1H), 7.65-7.58 (m, 1H), 7.55-7.46 (m, 1H), 7.32 (t, J=8.9 Hz, 1H), 7.28 (s, 1H), 7.04-6.95 (m, 3H), 6.82 (d, J=15.2 Hz, 1H), 5.02 (dd, J=12.3, 5.0 Hz, 1H), 4.13 (s, 3H), 4.03 (d, J=7.0 Hz, 2H), 3.33 (d, J=8.4 Hz, 8H), 3.25-3.08 (m, 2H), 2.84-2.80 (m, 1H), 2.74-2.65 (m, 2H), 2.48 (s, 2H), 2.12-2.03 (m, 1H), 1.69 (s, 4H). HRMS (ESI) m/z: calcd for C41H41ClFN9O7+ [M+H]+, 825.2802; found, 825.2800.
Referring to the method of example 1, the target compound (SIAIS249049) was prepared by using Dacomitinib derivative A and intermediate LM (SIAIS151027). (yellow solid, 8.3 mg, yield 47%)1H NMR (500 MHz, MeOD) δ 9.26 (s, 1H), 8.75 (s, 1H), 7.93 (dd, J=6.6, 2.6 Hz, 1H), 7.68-7.63 (m, 1H), 7.53 (dd, J=8.5, 7.2 Hz, 1H), 7.37 (t, J=8.9 Hz, 1H), 7.33 (s, 1H), 7.07-7.02 (m, 3H), 6.88 (d, J=15.2 Hz, 1H), 5.06 (dd, J=12.6, 5.5 Hz, 1H), 4.18 (s, 3H), 4.10 (d, J=7.2 Hz, 2H), 3.92-3.32 (m, 8H), 3.18 (d, J=12.6 Hz, 2H), 2.89-2.85 (m, 1H), 2.76-2.67 (m, 2H), 2.48 (t, J=7.2 Hz, 2H), 2.13-2.08 (m, 1H), 1.74-1.65 (m, 4H), 1.49-1.46 (m, 2H). HRMS (ESI) m/z: calcd for C42H43ClFN9O7+ [M+H]+, 839.2958; found, 839.2955.
Referring to the method of example 1, the target compound (SIAIS262015) was prepared by using Dacomitinib derivative A and intermediate LM (SIAIS151086). (yellow solid, 8.9 mg, yield 50%)1H NMR (500 MHz, MeOD) δ 9.27 (s, 1H), 8.75 (s, 1H), 7.93 (dd, J=6.6, 2.6 Hz, 1H), 7.68-7.63 (m, 1H), 7.54 (dd, J=8.6, 7.1 Hz, 1H), 7.38 (t, J=8.9 Hz, 1H), 7.32 (s, 1H), 7.09-7.01 (m, 3H), 6.87 (d, J=15.2 Hz, 1H), 5.06 (dd, J=12.5, 5.5 Hz, 1H), 4.18 (s, 3H), 4.07 (d, J=7.0 Hz, 2H), 3.38-3.31 (m, 8H), 2.88-2.84 (m, 1H), 2.77-2.68 (m, 2H), 2.44 (t, J=7.5 Hz, 2H), 2.16-2.07 (m, 1H), 1.68-1.63 (m, 4H), 1.51-1.42 (m, 4H), 1.33-1.28 (m, 2H). HRMS (ESI) m/z: calcd for C43H45ClFN9O7+ [M+H]+, 853.3115; found, 853.3111.
Referring to the method of example 1, the target compound (SIAIS249056) was prepared by using Dacomitinib derivative A and intermediate LM (SIAIS1204057). (yellow solid, 7.1 mg, yield 44%)1H NMR (500 MHz, MeOD) δ 9.28 (d, J=6.8 Hz, 1H), 8.76 (s, 1H), 7.95-7.91 (m, 1H), 7.68-7.63 (m, 1H), 7.43-7.31 (m, 3H), 7.15 (dd, J=7.5, 4.7 Hz, 1H), 7.07-7.02 (m, 1H), 6.93-6.82 (m, 2H), 5.21 (dd, J=13.3, 5.2 Hz, 1H), 4.82-4.29 (m, 4H), 4.19 (s, 3H), 4.12 (t, J=7.7 Hz, 2H), 3.73-3.32 (m, 8H), 3.23-3.08 (m, 2H), 2.98-2.89 (m, 1H), 2.80 (d, J=18.4 Hz, 1H), 2.58-2.44 (m, 1H), 2.21 (d, J=10.3 Hz, 1H). HRMS (ESI) m/z: calcd for C38H37ClFN9O6+ [M+H]+, 769.2539; found, 769.2535.
Referring to the method of example 1, the target compound (SIAIS249057) was prepared by using Dacomitinib derivative A and intermediate LM (SIAIS1204085). (yellow solid, 8.2 mg, yield 49%)1H NMR (500 MHz, MeOD) δ 9.27 (d, J=5.4 Hz, 1H), 8.75 (s, 1H), 7.93 (dd, J=6.6, 2.5 Hz, 1H), 7.67-7.63 (m, 1H), 7.53 (dd, J=9.7, 5.7 Hz, 1H), 7.47 (d, J=7.4 Hz, 1H), 7.37 (t, J=8.9 Hz, 1H), 7.34 (d, J=11.2 Hz, 1H), 7.28 (d, J=7.8 Hz, 1H), 7.08-6.98 (m, 1H), 6.85 (d, J=15.2 Hz, 1H), 5.17 (d, J=8.6 Hz, 1H), 4.74-4.46 (m, 2H), 4.19 (s, 3H), 4.03 (d, J=6.5 Hz, 2H), 3.62 (d, J=45.4 Hz, 2H), 3.46 (t, J=6.0 Hz, 2H), 3.34-3.31 (m, 8H), 3.14 (d, J=29.1 Hz, 2H), 2.98-2.84 (m, 1H), 2.79 (d, J=15.3 Hz, 1H), 2.68-2.47 (m, 2H), 2.29-2.17 (m, 1H), 2.05 (d, J=6.0 Hz, 1H). HRMS (ESI) m/z: calcd for C40H41ClFN9O6+ [M+H]+, 797.2852; found, 797.2851.
Referring to the method of example 1, the target compound (SIAIS249058) was prepared by using Dacomitinib derivative A and intermediate LM (SIAIS1210133). (yellow solid, 8.5 mg, yield 50%)1H NMR (500 MHz, MeOD) δ 9.29 (s, 1H), 8.76 (s, 1H), 7.93 (dd, J=6.6, 2.6 Hz, 1H), 7.66 (ddd, J=8.9, 4.1, 2.7 Hz, 1H), 7.58 (d, J=4.4 Hz, 2H), 7.38 (dd, J=12.1, 5.7 Hz, 2H), 7.34 (s, 1H), 7.06 (dt, J=14.3, 7.1 Hz, 1H), 6.88 (d, J=15.2 Hz, 1H), 5.20 (dd, J=13.1, 5.0 Hz, 1H), 4.62 (d, J=17.2 Hz, 1H), 4.55 (d, J=17.2 Hz, 1H), 4.19 (s, 3H), 4.09 (d, J=7.1 Hz, 2H), 3.60 (s, 2H), 3.44 (t, J=6.9 Hz, 2H), 3.31 (d, J=1.6 Hz, 8H), 2.97-2.86 (m, 1H), 2.80 (d, J=17.5 Hz, 1H), 2.54 (d, J=8.6 Hz, 2H), 2.22 (d, J=10.6 Hz, 1H), 1.87-1.71 (m, 4H). HRMS (ESI) m/z: calcd for C41H43ClFN9O6+ [M+H]+, 811.3009; found, 811.3007.
Referring to the method of example 1, the target compound (SIAIS249059) was prepared by using Dacomitinib derivative A and intermediate LM (SIAIS1204061). (yellow solid, 8.7 mg, yield 50%)1H NMR (500 MHz, MeOD) δ 9.27 (s, 1H), 8.75 (s, 1H), 7.93 (dd, J=6.6, 2.5 Hz, 1H), 7.68-7.57 (m, 3H), 7.44 (d, J=7.4 Hz, 1H), 7.36 (dd, J=17.4, 8.5 Hz, 2H), 7.12-7.02 (m, 1H), 6.89 (d, J=15.2 Hz, 1H), 5.20 (dd, J=13.2, 4.8 Hz, 1H), 4.64 (d, J=17.2 Hz, 1H), 4.57 (d, J=17.2 Hz, 1H), 4.18 (s, 3H), 4.10 (d, J=7.0 Hz, 2H), 3.63 (d, J=37.0 Hz, 2H), 3.42 (t, J=7.3 Hz, 2H), 3.31-3.11 (m, 8H), 2.98-2.76 (m, 2H), 2.60-2.44 (m, 3H), 2.27-2.17 (m, 1H), 1.88-1.63 (m, 4H), 1.58-1.45 (m, 2H). HRMS (ESI) m/z: calcd for C42H45ClFN9O6+ [M+H]+, 825.3165; found, 825.3163.
Referring to the method of example 1, the target compound (SIAIS249060) was prepared by using Dacomitinib derivative A and intermediate LM (SIAIS1204063). (yellow solid, 8.3 mg, yield 47%)1H NMR (500 MHz, MeOD) δ 9.28 (s, 1H), 8.75 (s, 1H), 7.93 (dd, J=6.6, 2.6 Hz, 1H), 7.68-7.64 (m, 1H), 7.40 (dt, J=17.8, 8.3 Hz, 2H), 7.33 (s, 1H), 7.26 (d, J=7.5 Hz, 1H), 7.09-7.02 (m, 2H), 6.88 (d, J=15.2 Hz, 1H), 5.17 (dd, J=13.3, 5.1 Hz, 1H), 4.43 (d, J=17.0 Hz, 1H), 4.37 (d, J=17.0 Hz, 1H), 4.09 (d, J=7.2 Hz, 2H), 3.57 (s, 2H), 3.31 (d, J=1.6 Hz, 8H), 2.95-2.88 (m, 1H), 2.83-2.75 (m, 1H), 2.59-2.38 (m, 3H), 2.23-2.18 (m, 1H), 1.69-1.64 (m, 4H), 1.52-1.39 (m, 4H). HRMS (ESI) m/z: calcd for C43H47ClFN9O6+ [M+H]+, 839.3322; found, 839.3322.
Referring to the method of example 1, the target compound (SIAIS249034) was prepared by using Dacomitinib derivative A and intermediate LM (SIAIS151045). (yellow solid, 8.2 mg, yield 49%)1H NMR (500 MHz, DMSO) δ 11.13 (s, 1H), 10.13 (s, 1H), 9.10 (s, 1H), 8.80 (s, 1H), 8.02 (d, J=4.9 Hz, 1H), 7.82-7.77 (m, 2H), 7.74-7.62 (m, 3H), 7.52 (t, J=9.0 Hz, 1H), 7.38 (s, 1H), 6.94 (s, 1H), 6.80 (d, J=15.8 Hz, 1H), 5.14-5.09 (m, 1H), 4.35 (s, 2H), 4.07 (s, 3H), 3.55-3.50 (m, 4H), 3.25-3.11 (m, 4H), 2.92-2.86 (m, 2H), 2.60 (d, J=17.4 Hz, 2H), 2.10-1.96 (m, 2H). HRMS (ESI) m/z: calcd for C38H35ClFN8O7S+ [M+H]+, 801.2016; found, 801.2013.
Referring to the method of example 1, the target compound (SIAIS249035) was prepared by using Dacomitinib derivative A and intermediate LM (SIAIS151138). (yellow solid, 9.1 mg, yield 53%)1H NMR (500 MHz, DMSO) δ 8.43 (s, 1H), 7.94 (s, 1H), 7.12 (s, 1H), 6.92 (d, J=17.4 Hz, 4H), 6.83 (d, J=25.9 Hz, 2H), 6.56 (t, J=8.8 Hz, 1H), 6.50 (s, 1H), 6.31-6.20 (m, 1H), 6.04 (d, J=15.5 Hz, 1H), 4.35-4.29 (m, 1H), 3.37 (s, 3H), 3.26 (s, 2H), 2.62-2.55 (m, 8H), 2.09-2.05 (m, 4H), 1.96-1.91 (m, 4H). HRMS (ESI) m/z: calcd for C39H37ClFN8O7S+ [M+H]+, 815.2173; found, 815.2170.
Referring to the method of example 1, the target compound (SIAIS249036) was prepared by using Dacomitinib derivative A and intermediate LM (SIAIS151139B). (yellow solid, 7.4 mg, yield 42%)1H NMR (500 MHz, DMSO) δ 11.61 (s, 1H), 11.12 (s, 1H), 10.16 (s, 1H), 9.13 (s, 1H), 8.84 (s, 1H), 8.00 (dd, J=6.6, 2.3 Hz, 1H), 7.86 (d, J=8.2 Hz, 1H), 7.80 (t, J=7.7 Hz, 1H), 7.72-7.66 (m, 1H), 7.64 (d, J=7.2 Hz, 1H), 7.53 (t, J=9.1 Hz, 1H), 7.41 (s, 1H), 6.96 (dd, J=14.7, 7.4 Hz, 1H), 6.80 (d, J=15.3 Hz, 1H), 5.11 (dd, J=12.9, 5.4 Hz, 1H), 4.07 (s, 3H), 3.99 (s, 2H), 3.16-3.11 (m, 8H), 2.99-2.83 (m, 2H), 2.65-2.53 (m, 4H), 2.07-1.96 (m, 2H), 1.91 (s, 2H). HRMS (ESI) m/z: calcd for C40H39ClFN8O7S+ [M+H]+, 829.2329; found, 829.2329.
Referring to the method of example 1, the target compound (SIAIS249037) was prepared by using Dacomitinib derivative A and intermediate LM (SIAIS151140B). (yellow solid, 8.5 mg, yield 48%)1H NMR (500 MHz, DMSO) δ 11.75 (s, 1H), 11.12 (s, 1H), 10.16 (s, 1H), 9.13 (s, 1H), 8.84 (s, 1H), 8.00 (dd, J=6.7, 2.4 Hz, 1H), 7.82-7.73 (m, 2H), 7.73-7.66 (m, 1H), 7.63 (d, J=6.9 Hz, 1H), 7.53 (t, J=9.0 Hz, 1H), 7.43 (s, 1H), 7.03-6.93 (m, 1H), 6.79 (d, J=15.4 Hz, 1H), 5.11 (dd, J=12.8, 5.4 Hz, 1H), 4.07 (s, 3H), 3.98 (s, 2H), 3.27-3.00 (m, 8H), 2.95-2.83 (m, 2H), 2.65-2.55 (m, 2H), 2.44 (s, 2H), 2.09-1.95 (m, 2H), 1.70 (s, 4H). HRMS (ESI) m/z: calcd for C41H41ClFN8O7S+ [M+H]+, 843.2486; found, 843.2483.
Referring to the method of example 1, the target compound (SIAIS249038) was prepared by using Dacomitinib derivative A and intermediate LM (SIAIS151141B). (yellow solid, 8.9 mg, yield 49%)1H NMR (500 MHz, DMSO) δ 11.62 (s, 1H), 11.12 (s, 1H), 10.15 (s, 1H), 9.12 (s, 1H), 8.83 (s, 1H), 8.00 (dd, J=6.8, 2.4 Hz, 1H), 7.81-7.73 (m, 2H), 7.73-7.66 (m, 1H), 7.63 (d, J=7.0 Hz, 1H), 7.52 (t, J=9.1 Hz, 1H), 7.41 (s, 1H), 6.98-6.84 (m, 1H), 6.79 (d, J=15.5 Hz, 1H), 5.11 (dd, J=12.9, 5.4 Hz, 1H), 4.07 (s, 3H), 3.99 (s, 2H), 3.16-3.09 (m, 8H), 2.97-2.82 (m, 2H), 2.60-2.53 (m, 2H), 2.38 (t, J=7.2 Hz, 2H), 2.04-1.99 (m, 2H), 1.71-1.66 (m, 2H), 1.55 (d, J=7.2 Hz, 2H), 1.49-1.44 (m, 2H). HRMS (ESI) m/z: calcd for C42H43ClFN8O7S+ [M+H]+, 857.2642; found, 857.2640.
Referring to the method of example 1, the target compound (SIAIS249039) was prepared by using Dacomitinib derivative A and intermediate LM (SIAIS151142B). (yellow solid, 9.1 mg, yield 50%)1H NMR (500 MHz, DMSO) δ 11.79-11.41 (m, 1H), 11.12 (s, 1H), 10.12 (s, 1H), 9.09 (s, 1H), 8.79 (s, 1H), 8.03 (s, 1H), 7.81-7.70 (m, 3H), 7.63 (d, J=7.0 Hz, 1H), 7.51 (t, J=9.0 Hz, 1H), 7.41 (s, 1H), 6.94 (s, 1H), 6.78 (d, J=14.9 Hz, 1H), 5.11 (dd, J=12.9, 5.4 Hz, 1H), 4.06 (s, 3H), 3.97 (s, 2H), 3.14-3.08 (m, 8H), 2.94-2.83 (m, 2H), 2.66-2.54 (m, 2H), 2.36 (t, J=7.0 Hz, 2H), 2.05-1.99 (m, 2H), 1.67 (dd, J=14.6, 7.2 Hz, 2H), 1.54-1.42 (m, 4H), 1.36-1.32 (m, 2H). HRMS (ESI) m/z: calcd for C43H45ClFN8O7S+ [M+H]+, 871.2799; found, 871.2797.
Referring to the method of example 1, the target compound (SIAIS219192) was prepared by using Dacomitinib derivative A and intermediate LM (SIAIS1204137). (yellow solid, 9.5 mg, yield 54%)1H NMR (500 MHz, MeOD) δ 9.20 (s, 1H), 8.76 (d, J=4.8 Hz, 1H), 7.94 (dd, J=6.6, 2.6 Hz, 1H), 7.72 (dd, J=12.5, 7.9 Hz, 2H), 7.69-7.64 (m, 1H), 7.55 (t, J=5.4 Hz, 1H), 7.37 (t, J=8.9 Hz, 1H), 7.32 (s, 1H), 7.17-7.10 (m, 1H), 6.88 (d, J=15.2 Hz, 1H), 5.11 (dd, J=12.8, 5.5 Hz, 1H), 4.40-4.36 (m, 2H), 4.22-4.07 (m, 8H), 3.89-3.84 (m, 2H), 3.66 (s, 2H), 3.19-3.17 (m, 2H), 2.91-2.85 (m, 1H), 2.75-2.70 (m, 2H), 2.16-2.09 (m, 1H). HRMS (ESI) m/z: calcd for C40H39ClFN8O8S+ [M+H]+, 845.2279; found, 845.2275.
Referring to the method of example 1, the target compound (SIAIS262005) was prepared by using Dacomitinib derivative A and intermediate LM (SIAIS1204139). (yellow solid, 9.1 mg, yield 49%)1H NMR (500 MHz, DMSO) δ 11.12 (s, 1H), 10.12 (s, 1H), 9.11 (s, 1H), 8.83 (s, 1H), 8.01 (s, 1H), 7.80-7.74 (m, 2H), 7.70 (s, 1H), 7.61 (d, J=6.0 Hz, 1H), 7.53 (s, 1H), 7.36 (s, 1H), 6.94 (s, 1H), 6.79 (d, J=14.0 Hz, 1H), 5.12 (dd, J=12.9, 5.4 Hz, 1H), 4.22 (d, J=15.0 Hz, 2H), 4.07 (s, 3H), 4.02 (s, 2H), 3.73 (t, J=6.3 Hz, 2H), 3.55-3.50 (m, 10H), 3.09 (s, 4H), 2.92-2.86 (m, 1H), 2.64-2.55 (m, 2H), 2.09-2.01 (m, 1H). HRMS (ESI) m/z: calcd for C42H43ClFN8O9S+ [M+H]+, 889.2541; found, 889.2543.
Referring to the method of example 1, the target compound (SIAIS262006) was prepared by using Dacomitinib derivative A and intermediate LM (SIAIS1204141). (yellow solid, 9.7 mg, yield 49%)1H NMR (500 MHz, DMSO) δ 11.44-11.23 (m, 1H), 11.12 (s, 1H), 10.13 (s, 1H), 9.10 (s, 1H), 8.82 (s, 1H), 8.01 (s, 1H), 7.80-7.74 (m, 2H), 7.69 (s, 1H), 7.62 (dd, J=5.9, 2.0 Hz, 1H), 7.52 (t, J=8.9 Hz, 1H), 7.37 (s, 1H), 6.93 (s, 1H), 6.79 (d, J=14.8 Hz, 1H), 5.11 (dd, J=12.9, 5.4 Hz, 1H), 4.20 (d, J=16.9 Hz, 2H), 4.07 (s, 3H), 4.01 (s, 2H), 3.71 (t, J=6.3 Hz, 2H), 3.58-3.53 (m, 8H), 3.34 (t, J=6.3 Hz, 6H), 3.07 (s, 4H), 2.93-2.84 (m, 1H), 2.66-2.57 (m, 2H), 2.09-2.02 (m, 1H). HRMS (ESI) m/z: calcd for C44H47ClFN8O10S+ [M+H]+, 933.2803; found, 933.2800.
Referring to the method of example 1, the target compound (SIAIS262007) was prepared by using Dacomitinib derivative A and intermediate LM (SIAIS1204147). (yellow solid, 10.1 mg, yield 49%) 1H NMR (500 MHz, DMSO) δ 11.72-11.32 (m, 1H), 11.12 (s, 1H), 10.16 (s, 1H), 9.14 (s, 1H), 8.85 (s, 1H), 8.00 (s, 1H), 7.79-7.74 (m, 2H), 7.68 (s, 1H), 7.62 (dd, J=5.5, 2.4 Hz, 1H), 7.53 (t, J=9.1 Hz, 1H), 7.39 (s, 1H), 6.96 (s, 1H), 6.80 (d, J=15.1 Hz, 1H), 5.11 (dd, J=12.9, 5.4 Hz, 1H), 4.20 (d, J=20.5 Hz, 2H), 4.07 (s, 3H), 4.01 (s, 2H), 3.70 (t, J=6.3 Hz, 2H), 3.59-3.50 (m, 14H), 3.34 (t, J=6.3 Hz, 4H), 3.03 (d, J=50.8 Hz, 4H), 2.92-2.88 (m, 1H), 2.64-2.53 (m, 2H), 2.09-2.01 (m, 1H). HRMS (ESI) m/z: calcd for C46H51ClFN8O1S+ [M+H]+, 977.3065; found, 977.3061.
Referring to the method of example 1, the target compound (SIAIS262008) was prepared by using Dacomitinib derivative A and intermediate LM (SIAIS1204149). (yellow solid, 11.5 mg, yield 54%) 1H NMR (500 MHz, DMSO) δ 11.89 (s, 1H), 11.12 (s, 1H), 10.17 (s, 1H), 9.15 (s, 1H), 8.86 (s, 1H), 7.99 (dd, J=6.8, 2.5 Hz, 1H), 7.79-7.74 (m, 2H), 7.69-7.66 (m, 1H), 7.65-7.60 (m, 1H), 7.53 (td, J=9.0, 3.2 Hz, 1H), 7.45 (s, 1H), 7.02-6.94 (m, 1H), 6.80 (d, J=15.3 Hz, 1H), 5.11 (dd, J=12.9, 5.4 Hz, 1H), 4.21 (s, 2H), 4.07 (s, 3H), 4.00 (d, J=9.4 Hz, 2H), 3.70 (t, J=6.3 Hz, 2H), 3.61-3.51 (m, 18H), 3.33 (t, J=6.3 Hz, 4H), 3.06 (d, J=41.3 Hz, 4H), 2.91-2.86 (m, 1H), 2.62-2.52 (m, 2H), 2.08-1.99 (m, 1H). HRMS (ESI) m/z: calcd for C48H55ClFN8O12S+ [M+H]+, 1021.3327; found, 1021.3324.
Referring to the method of example 1, the target compound (SIAIS219185) was prepared by using Dacomitinib derivative A and intermediate LM (SIAIS171090). (yellow solid, 8.1 mg, yield 49%)1H NMR (500 MHz, MeOD) δ 9.27 (s, 1H), 8.75 (s, 1H), 7.93 (dd, J=6.6, 2.6 Hz, 1H), 7.82-7.77 (m, 2H), 7.74 (d, J=7.5 Hz, 1H), 7.68-7.62 (m, 1H), 7.56 (t, J=7.7 Hz, 1H), 7.44 (d, J=8.1 Hz, 1H), 7.37 (t, J=8.9 Hz, 1H), 7.32 (s, 1H), 7.05-7.00 (m, 1H), 6.86 (d, J=15.2 Hz, 1H), 5.18 (dd, J=13.3, 5.2 Hz, 1H), 4.56 (d, J=17.5 Hz, 1H), 4.49 (d, J=17.4 Hz, 1H), 4.18 (s, 3H), 4.16-4.12 (m, 2H), 4.08 (d, J=5.9 Hz, 2H), 4.03 (d, J=5.4 Hz, 2H), 3.77-3.53 (m, 6H), 2.94-2.87 (m, 1H), 2.82-2.76 (m, 1H), 2.58-2.52 (m, 1H), 2.21-2.17 (m, 1H). HRMS (ESI) m/z: calcd for C38H37ClFN8O6S+ [M+H]+, 787.2224; found, 787.2221.
Referring to the method of example 1, the target compound (SIAIS219186) was prepared by using Dacomitinib derivative A and intermediate LM (SIAIS171086). (yellow solid, 8.5 mg, yield 51%)1H NMR (500 MHz, MeOD) δ 9.27 (s, 1H), 8.75 (s, 1H), 7.93 (dd, J=6.6, 2.6 Hz, 1H), 7.74-7.68 (m, 2H), 7.69-7.64 (m, 1H), 7.56 (t, J=7.7 Hz, 1H), 7.37 (t, J=8.9 Hz, 1H), 7.32 (s, 1H), 7.08-7.00 (m, 1H), 6.84 (d, J=15.2 Hz, 1H), 5.19 (dd, J=13.4, 5.1 Hz, 1H), 4.49 (d, J=17.4 Hz, 1H), 4.43 (d, J=17.4 Hz, 1H), 4.18 (s, 3H), 4.02 (d, J=6.8 Hz, 2H), 3.96-3.32 (m, 8H), 3.17 (s, 2H), 2.95-2.90 (m, 1H), 2.83-2.76 (m, 3H), 2.56-2.52 (m, 1H), 2.21-2.18 (m, 1H). HRMS (ESI) m/z: calcd for C39H39ClFN8O6S+ [M+H]+, 801.2380; found, 801.2381.
Referring to the method of example 1, the target compound (SIAIS219187) was prepared by using Dacomitinib derivative A and intermediate LM (SIAIS171089). (yellow solid, 9.0 mg, yield 53%)1H NMR (500 MHz, MeOD) δ 9.29 (s, 1H), 8.76 (s, 1H), 7.94 (dd, J=6.6, 2.6 Hz, 1H), 7.72 (d, J=7.7 Hz, 1H), 7.70-7.64 (m, 2H), 7.55 (t, J=7.7 Hz, 1H), 7.38 (t, J=8.9 Hz, 1H), 7.33 (s, 1H), 7.07-7.02 (m, 1H), 6.87 (d, J=15.2 Hz, 1H), 5.18 (dd, J=13.3, 5.1 Hz, 1H), 4.49 (d, J=17.4 Hz, 1H), 4.46-4.40 (m, 1H), 4.19 (s, 3H), 4.06 (d, J=7.0 Hz, 2H), 3.72-3.62 (m, 2H), 3.59-3.49 (m, 2H), 3.25-3.08 (m, 5H), 2.94-2.86 (m, 2H), 2.83-2.74 (m, 1H), 2.62-2.50 (m, 3H), 2.21-2.14 (m, 1H), 1.97-1.93 (m, 2H). HRMS (ESI) m/z: calcd for C40H41ClFN8O6S+ [M+H]+, 815.2537; found, 815.2535.
Referring to the method of example 1, the target compound (SIAIS219188) was prepared by using Dacomitinib derivative A and intermediate LM (SIAIS171079). (yellow solid, 8.8 mg, yield 51%)1H NMR (500 MHz, MeOD) δ 9.28 (s, 1H), 8.75 (s, 1H), 7.94 (dd, J=6.6, 2.6 Hz, 1H), 7.68-7.63 (m, 3H), 7.53 (t, J=7.7 Hz, 1H), 7.38 (t, J=8.9 Hz, 1H), 7.32 (s, 1H), 7.09-7.03 (m, 1H), 6.85 (d, J=15.2 Hz, 1H), 5.17 (dd, J=13.4, 5.1 Hz, 1H), 4.48 (d, J=17.4 Hz, 1H), 4.43 (d, J=17.4 Hz, 1H), 4.18 (s, 3H), 4.02 (s, 2H), 3.15-3.10 (m, 8H), 2.96-2.85 (m, 1H), 2.82-2.76 (m, 1H), 2.57-2.52 (m, 1H), 2.45 (t, J=7.1 Hz, 2H), 2.22-2.15 (m, 2H), 2.04 (s, 1H), 1.77-1.68 (m, 4H). HRMS (ESI) m/z: calcd for C41H43ClFN8O6S+ [M+H]+, 829.2693; found, 829.2690.
Referring to the method of example 1, the target compound (SIAIS219189) was prepared by using Dacomitinib derivative A and intermediate LM (SIAIS171091). (yellow solid, 9.1 mg, yield 51%)1H NMR (500 MHz, MeOD) δ 9.26 (s, 1H), 8.75 (s, 1H), 7.93 (dd, J=6.6, 2.6 Hz, 1H), 7.68-7.63 (m, 3H), 7.53 (t, J=7.7 Hz, 1H), 7.38 (t, J=8.9 Hz, 1H), 7.32 (s, 1H), 7.09-7.03 (m, 1H), 6.87 (d, J=15.3 Hz, 1H), 5.17 (dd, J=13.3, 5.2 Hz, 1H), 4.50-4.44 (m, 1H), 4.42 (d, J=17.3 Hz, 1H), 4.18 (s, 3H), 4.05 (d, J=7.2 Hz, 2H), 3.34-3.30 (m, 8H), 3.09-3.05 (m, 2H), 2.95-2.88 (m, 1H), 2.79-2.74 (m, 1H), 2.57-2.52 (m, 1H), 2.38 (t, J=7.3 Hz, 2H), 2.22-2.14 (m, 1H), 1.66-1.62 (m, 4H), 1.53-1.49 (m, 2H). HRMS (ESI) m/z: calcd for C42H45ClFN8O6S+ [M+H]+, 843.2850; found, 843.2852.
Referring to the method of example 1, the target compound (SIAIS219190) was prepared by using Dacomitinib derivative A and intermediate LM (SIAIS171092). (yellow solid, 8.9 mg, yield 49%)1H NMR (500 MHz, MeOD) δ 9.26 (s, 1H), 8.74 (s, 1H), 7.93 (dd, J=6.6, 2.6 Hz, 1H), 7.67-7.62 (m, 3H), 7.52 (t, J=7.7 Hz, 1H), 7.37 (t, J=8.9 Hz, 1H), 7.32 (s, 1H), 7.05 (dd, J=14.9, 7.4 Hz, 1H), 6.86 (d, J=15.2 Hz, 1H), 5.17 (dd, J=13.4, 5.2 Hz, 1H), 4.49-4.43 (m, 1H), 4.40 (d, J=17.3 Hz, 1H), 4.18 (s, 3H), 4.04 (d, J=7.0 Hz, 2H), 3.31 (d, J=1.6 Hz, 8H), 3.10-3.02 (m, 2H), 2.92-2.88 (m, 1H), 2.82-2.74 (m, 1H), 2.57-2.52 (m, 1H), 2.39 (t, J=7.5 Hz, 2H), 2.22-2.14 (m, 1H), 1.71-1.63 (m, 2H), 1.59-1.55 (m, 2H), 1.49-1.44 (m, 2H), 1.39-1.34 (m, 2H). HRMS (ESI) m/z: calcd for C43H47ClFN8O6S+ [M+H]+, 857.3006; found, 857.3003.
Referring to the method of example 1, the target compound (SIAIS219193) was prepared by using Dacomitinib derivative A and intermediate LM (SIAIS1213129). (yellow solid, 9.1 mg, yield 52%)1H NMR (500 MHz, MeOD) δ 9.29 (s, 1H), 8.76 (s, 1H), 7.94 (dd, J=6.6, 2.6 Hz, 1H), 7.72 (d, J=7.7 Hz, 1H), 7.70-7.61 (m, 2H), 7.52 (dd, J=10.0, 5.3 Hz, 1H), 7.38 (t, J=8.9 Hz, 1H), 7.32 (s, 1H), 7.07 (dt, J=14.8, 7.2 Hz, 1H), 6.86 (d, J=15.2 Hz, 1H), 5.17 (dd, J=13.1, 5.1 Hz, 1H), 4.51 (d, J=17.5 Hz, 1H), 4.47 (d, J=8.0 Hz, 1H), 4.21 (d, J=5.0 Hz, 2H), 4.18 (s, 3H), 4.09-4.05 (m, 2H), 3.74 (dd, J=10.6, 4.8 Hz, 2H), 3.35-3.31 (m, 8H), 3.26 (dd, J=13.1, 6.9 Hz, 2H), 2.91-2.86 (m, 1H), 2.83-2.76 (m, 1H), 2.58-2.54 (m, 1H), 2.25-2.17 (m, 1H). HRMS (ESI) m/z: calcd for C40H41ClFN8O7S+ [M+H]+, 831.2486; found, 831.2482.
Referring to the method of example 1, the target compound (SIAIS262001) was prepared by using Dacomitinib derivative A and intermediate LM (SIAIS1213131). (yellow solid, 9.2 mg, yield 50%)1H NMR (500 MHz, DMSO) δ 11.50 (s, 1H), 10.99 (s, 1H), 10.15 (s, 1H), 9.13 (s, 1H), 8.85 (s, 1H), 8.00 (d, J=6.8 Hz, 1H), 7.69 (d, J=7.7 Hz, 2H), 7.59-7.51 (m, 3H), 7.39 (s, 1H), 7.00-6.92 (m, 1H), 6.80 (d, J=15.4 Hz, 1H), 5.13 (dd, J=13.3, 5.2 Hz, 1H), 4.37 (d, J=17.4 Hz, 1H), 4.24 (d, J=17.5 Hz, 1H), 4.20 (d, J=9.9 Hz, 2H), 4.07 (s, 3H), 4.01 (s, 2H), 3.63 (t, J=6.4 Hz, 2H), 3.52 (d, J=34.2 Hz, 8H), 3.26 (t, J=6.1 Hz, 2H), 3.17-2.92 (m, 4H), 2.93-2.88 (m, 1H), 2.65-2.58 (m, 1H), 2.47-2.43 (m, 1H), 2.05-1.99 (m, 1H). HRMS (ESI) m/z: calcd for C42H45ClFN8O8S+ [M+H]+, 875.2748; found, 875.2744.
Referring to the method of example 1, the target compound (SIAIS262002) was prepared by using Dacomitinib derivative A and intermediate LM (SIAIS1213133). (yellow solid, 10.2 mg, yield 53%)1H NMR (500 MHz, DMSO) δ 10.99 (s, 1H), 10.15 (s, 1H), 9.13 (s, 1H), 8.85 (s, 1H), 8.00 (dd, J=6.8, 2.5 Hz, 1H), 7.71-7.65 (m, 2H), 7.59-7.51 (m, 3H), 7.41 (s, 1H), 7.03-6.93 (m, 1H), 6.80 (d, J=15.4 Hz, 1H), 5.12 (dd, J=13.3, 5.1 Hz, 1H), 4.37 (d, J=17.4 Hz, 1H), 4.25-4.21 (m, 1H), 4.21 (s, 2H), 4.07 (s, 3H), 4.02-3.95 (m, 2H), 3.61 (d, J=6.3 Hz, 2H), 3.57-3.52 (m, 8H), 3.25 (t, J=6.0 Hz, 2H), 3.05 (s, 4H), 2.95-2.88 (m, 1H), 2.60 (t, J=15.9 Hz, 1H), 2.48-2.42 (m, 1H), 2.05-1.98 (m, 1H). HRMS (ESI) m/z: calcd for C44H49ClFN8O9S+ [M+H]+, 919.3010; found, 919.3012.
Referring to the method of example 1, the target compound (SIAIS262003) was prepared by using Dacomitinib derivative A and intermediate LM (SIAIS1213135). (yellow solid, 10.3 mg, yield 51%)1H NMR (500 MHz, DMSO) δ 11.54 (s, 1H), 10.99 (s, 1H), 10.15 (s, 1H), 9.13 (s, 1H), 8.84 (s, 1H), 8.00 (d, J=7.0 Hz, 1H), 7.70-7.67 (m, 2H), 7.56 (ddd, J=18.6, 5.8, 2.8 Hz, 3H), 6.96 (dd, J=15.1, 7.0 Hz, 1H), 6.80 (d, J=15.3 Hz, 1H), 5.15-5.11 (m, 1H), 4.37 (d, J=12.5 Hz, 1H), 4.23 (d, J=13.4 Hz, 1H), 4.19 (d, J=14.0 Hz, 2H), 4.07 (s, 3H), 4.01 (s, 2H), 3.63-3.61 (m, 2H), 3.54-3.51 (m, 8H), 3.26-3.25 (m, 2H), 3.08 (s, 4H), 2.93-2.88 (m, 1H), 2.59 (d, J=16.1 Hz, 1H), 2.45-2.41 (m, 1H), 2.02-2.00 (m, 1H). HRMS (ESI) m/z: calcd for C46H53ClFN8O10S+ [M+H]+, 963.3272; found, 963.3270.
Referring to the method of example 1, the target compound (SIAIS262004) was prepared by using Dacomitinib derivative A and intermediate LM (SIAIS1213137). (yellow solid, 11.2 mg, yield 53%)1H NMR (500 MHz, DMSO) δ 11.25 (s, 1H), 10.99 (s, 1H), 10.18 (s, 1H), 9.16 (s, 1H), 8.87 (s, 1H), 8.02-7.96 (m, 1H), 7.70-7.66 (m, 2H), 7.55 (tdd, J=12.9, 7.0, 3.5 Hz, 3H), 7.45 (d, J=2.0 Hz, 1H), 6.99 (dt, J=14.4, 7.1 Hz, 1H), 6.80 (d, J=15.3 Hz, 1H), 5.12 (dd, J=13.3, 5.1 Hz, 1H), 4.37 (dd, J=17.4, 4.1 Hz, 1H), 4.26-4.22 (m, 1H), 4.23-4.16 (m, 2H), 4.08 (d, J=1.9 Hz, 3H), 4.01 (t, J=5.5 Hz, 2H), 3.73-3.69 (m, 1H), 3.69-3.64 (m, 1H), 3.64-3.60 (m, 2H), 3.58-3.52 (m, 10H), 3.28-3.24 (m, 2H), 3.23-2.95 (m, 4H), 2.91 (ddd, J=13.6, 11.1, 5.5 Hz, 1H), 2.65-2.56 (m, 1H), 2.44 (d, J=13.1 Hz, 1H), 2.03-1.98 (m, 1H). HRMS (ESI) m/z: calcd for C48H57ClFN8O11S+ [M+H]+, 1007.3535; found, 1007.3533.
Referring to the method of example 1, the target compound (SIAIS249045) was prepared by using Dacomitinib derivative A and intermediate LM (SIAIS151002). (white solid, 11.1 mg, yield 49%)1H NMR (500 MHz, MeOD) δ 9.84 (s, 1H), 9.27 (s, 1H), 8.76 (s, 1H), 7.92 (dd, J=6.6, 2.6 Hz, 1H), 7.68-7.63 (m, 1H), 7.53 (dt, J=16.8, 6.5 Hz, 5H), 7.37 (dd, J=15.8, 6.9 Hz, 2H), 7.08 (dd, J=14.9, 7.4 Hz, 1H), 6.89 (d, J=15.2 Hz, 1H), 4.65 (s, 1H), 4.61-4.55 (m, 2H), 4.51 (d, J=12.2 Hz, 2H), 4.43 (s, 1H), 4.18 (s, 3H), 4.12 (d, J=7.1 Hz, 2H), 3.90 (d, J=11.1 Hz, 1H), 3.83-3.72 (m, 7H), 3.62 (t, J=9.1 Hz, 7H), 2.62-2.51 (m, 8H), 2.26-2.22 (m, 1H), 2.09-2.05 (m, 1H), 1.34-1.29 (m, 2H), 1.04 (s, 9H). HRMS (ESI) m/z: calcd for C53H65ClFN10O9S+ [M+H]+, 1071.4324; found, 1071.4321.
Referring to the method of example 1, the target compound (SIAIS249041) was prepared by using Dacomitinib derivative A and intermediate LM (SIAIS074011). (white solid, 10.2 mg, yield 49%)1H NMR (500 MHz, MeOD) δ 9.83 (s, 1H), 9.27 (s, 1H), 8.76 (s, 1H), 7.93 (dd, J=6.6, 2.5 Hz, 1H), 7.68-7.63 (m, 1H), 7.54 (dt, J=8.2, 3.5 Hz, 5H), 7.38 (d, J=8.9 Hz, 1H), 7.12-7.05 (m, 1H), 6.90 (d, J=15.0 Hz, 1H), 4.62-4.48 (m, 6H), 4.39 (dd, J=15.8, 6.1 Hz, 2H), 4.19 (s, 3H), 4.12 (d, J=6.9 Hz, 2H), 3.89 (d, J=11.0 Hz, 1H), 3.80 (dd, J=10.9, 3.7 Hz, 1H), 3.69-3.55 (m, 3H), 2.78-2.48 (m, 10H), 2.29-2.20 (m, 2H), 2.12-1.97 (m, 2H), 1.03 (s, 9H). HRMS (ESI) m/z: calcd for C49H57ClF10O7S+ [M+H]+, 983.3799; found, 983.3795.
Referring to the method of example 1, the target compound (SIAIS249042) was prepared by using Dacomitinib derivative A and intermediate LM (SIAIS074013). (white solid, 10.8 mg, yield 51%)1H NMR (500 MHz, MeOD) δ 9.61 (s, 1H), 9.27 (s, 1H), 8.76 (s, 1H), 7.92 (dd, J=6.6, 2.6 Hz, 1H), 7.65 (ddd, J=8.8, 4.1, 2.6 Hz, 1H), 7.56-7.48 (m, 4H), 7.38 (t, J=8.9 Hz, 1H), 7.34 (s, 1H), 7.07 (dd, J=14.9, 7.4 Hz, 1H), 6.89 (d, J=15.3 Hz, 1H), 4.63 (s, 1H), 4.57 (dd, J=14.8, 6.0 Hz, 2H), 4.51 (d, J=12.2 Hz, 2H), 4.41 (d, J=15.7 Hz, 1H), 4.19 (s, 3H), 4.10 (d, J=7.2 Hz, 2H), 3.91 (d, J=11.0 Hz, 1H), 3.80 (dd, J=10.9, 3.8 Hz, 1H), 3.75-3.48 (m, 3H), 2.56 (s, 3H), 2.48 (d, J=3.1 Hz, 2H), 2.36-2.31 (m, 2H), 2.25-2.20 (m, 1H), 2.09-2.05 (m, 1H), 1.69-1.62 (m, 4H), 1.04 (s, 9H). HRMS (ESI) m/z: calcd for C51H61ClFN10O7S+ [M+H]+, 1011.4112; found, 1011.4110.
Referring to the method of example 1, the target compound (SIAIS249043) was prepared by using Dacomitinib derivative A and intermediate LM (SIAIS074015). (white solid, 10.8 mg, yield 49%)1H NMR (500 MHz, MeOD) δ 9.86 (s, 1H), 9.27 (s, 1H), 8.76 (s, 1H), 7.93 (dd, J=6.6, 2.6 Hz, 1H), 7.66 (ddd, J=8.9, 4.1, 2.7 Hz, 1H), 7.54 (dd, J=23.3, 8.2 Hz, 4H), 7.39 (d, J=8.9 Hz, 1H), 7.35 (d, J=6.7 Hz, 1H), 7.10-7.06 (m, 1H), 6.89 (d, J=15.2 Hz, 1H), 4.64 (s, 1H), 4.59-4.48 (m, 3H), 4.41 (d, J=15.7 Hz, 1H), 4.19 (s, 3H), 4.11 (d, J=7.1 Hz, 2H), 3.91 (d, J=11.0 Hz, 1H), 3.82-3.78 (m, 1H), 2.59 (s, 3H), 2.46 (t, J=7.5 Hz, 2H), 2.33-2.20 (m, 3H), 2.09-2.05 (m, 1H), 1.66-1.57 (m, 4H), 1.03 (s, 9H). HRMS (ESI) m/z: calcd for C53H65ClFN10O7S+ [M+H]+, 1039.4425; found, 1039.4423.
Referring to the method of example 1, the target compound (SIAIS262032) was prepared by using Dacomitinib derivative B and intermediate LM (SIAIS151141B). (yellow solid, 8.1 mg, yield 48%)1H NMR (500 MHz, MeOD) δ 9.27 (s, 1H), 8.75 (s, 1H), 7.93 (dd, J=6.5, 2.5 Hz, 1H), 7.76-7.69 (m, 2H), 7.67-7.63 (m, 1H), 7.63-7.57 (m, 1H), 7.38 (t, J=8.9 Hz, 1H), 7.32 (s, 1H), 7.05 (dd, J=14.8, 7.3 Hz, 1H), 6.87 (d, J=15.2 Hz, 1H), 5.13 (dd, J=12.7, 5.4 Hz, 1H), 4.18 (s, 3H), 4.07 (d, J=6.4 Hz, 2H), 3.92 (s, 2H), 3.77 (d, J=12.3 Hz, 2H), 3.60 (s, 2H), 3.41-3.36 (m, 5H), 3.22 (s, 2H), 3.15 (t, J=7.1 Hz, 2H), 2.87 (t, J=8.7 Hz, 1H), 2.80-2.68 (m, 2H), 2.55-2.42 (m, 4H), 2.22 (d, J=10.3 Hz, 2H), 2.18-2.10 (m, 1H), 1.85-1.76 (m, 2H), 1.74-1.64 (m, 2H), 1.58 (d, J=6.9 Hz, 2H). HRMS (ESI) m/z: calcd for C47H52ClFN9O7S+ [M+H]+, 940.3377; found, 940.3374.
Referring to the method of example 1, the target compound (SIAIS262033) was prepared by using Dacomitinib derivative B and intermediate LM (SIAIS151142B). (yellow solid, 8.4 mg, yield 49%)1H NMR (500 MHz, MeOD) δ 9.26 (s, 1H), 8.75 (s, 1H), 7.93 (dd, J=6.6, 2.6 Hz, 1H), 7.74-7.68 (m, 2H), 7.68-7.64 (m, 1H), 7.60 (dd, J=6.8, 1.0 Hz, 1H), 7.38 (t, J=8.9 Hz, 1H), 7.32 (s, 1H), 7.10-7.02 (m, 1H), 6.87 (d, J=15.1 Hz, 1H), 5.16-5.12 (m, 1H), 4.18 (d, J=6.0 Hz, 3H), 4.08 (d, J=6.4 Hz, 2H), 3.92 (s, 2H), 3.77 (d, J=11.9 Hz, 2H), 3.61 (s, 2H), 3.45-3.42 (m, 5H), 3.22 (s, 2H), 3.13 (t, J=7.1 Hz, 2H), 2.90-2.86 (m, 1H), 2.76-2.72 (m, 2H), 2.52-2.42 (m, 4H), 2.22 (d, J=12.3 Hz, 2H), 2.18-2.11 (m, 1H), 1.81-1.73 (m, 2H), 1.66-1.62 (m, 2H), 1.58-1.53 (m, 2H), 1.47-1.42 (m, 2H). HRMS (ESI) m/z: calcd for C48H54ClFN9O7S+ [M+H]+, 954.3534; found, 954.3532.
Referring to the method of example 1, the target compound (SIAIS262034) was prepared by using Dacomitinib derivative B and intermediate LM (SIAIS171090). (yellow solid, 6.5 mg, yield 42%)1H NMR (500 MHz, MeOD) δ 9.28 (s, 1H), 8.75 (s, 1H), 7.93 (dd, J=6.6, 2.6 Hz, 1H), 7.83 (d, J=7.7 Hz, 1H), 7.76 (d, J=7.5 Hz, 1H), 7.70-7.63 (m, 1H), 7.58 (t, J=7.7 Hz, 1H), 7.38 (t, J=8.9 Hz, 1H), 7.32 (s, 1H), 7.07 (dt, J=14.6, 7.1 Hz, 1H), 6.88 (d, J=15.3 Hz, 1H), 5.17 (dd, J=13.3, 5.1 Hz, 1H), 4.57 (d, J=17.6 Hz, 1H), 4.50 (d, J=11.6 Hz, 1H), 4.18 (s, 3H), 4.12-3.98 (m, 4H), 3.78 (d, J=11.7 Hz, 2H), 3.58 (s, 2H), 3.38 (d, J=65.5 Hz, 5H), 3.20 (d, J=27.5 Hz, 4H), 2.93-2.87 (m, 1H), 2.82-2.78 (m, 1H), 2.59-2.54 (m, 1H), 2.42 (s, 1H), 2.21-2.18 (m, 4H). HRMS (ESI) m/z: calcd for C43H46ClFN9O6S+ [M+H]+, 870.2959; found, 870.2955.
Referring to the method of example 1, the target compound (SIAIS262035) was prepared by using Dacomitinib derivative B and intermediate LM (SIAIS171091). (yellow solid, 8.3 mg, yield 50%)1H NMR (500 MHz, MeOD) δ 9.27 (s, 1H), 8.75 (s, 1H), 7.93 (dd, J=6.6, 2.6 Hz, 1H), 7.69-7.63 (m, 3H), 7.54 (t, J=7.6 Hz, 1H), 7.38 (t, J=8.9 Hz, 1H), 7.33 (s, 1H), 7.11-7.01 (m, 1H), 6.88 (d, J=15.1 Hz, 1H), 5.17 (dd, J=13.3, 5.1 Hz, 1H), 4.48 (d, J=17.3 Hz, 1H), 4.42 (d, J=17.4 Hz, 1H), 4.19 (s, 3H), 4.09 (s, 2H), 3.79 (d, J=10.9 Hz, 2H), 3.63 (d, J=12.1 Hz, 2H), 3.50-3.32 (m, 5H), 3.27-3.21 (m, 2H), 3.20-3.01 (m, 4H), 2.94-2.90 (m, 1H), 2.84-2.77 (m, 1H), 2.62-2.54 (m, 1H), 2.50 (d, J=10.6 Hz, 2H), 2.39 (t, J=7.2 Hz, 2H), 2.30-2.15 (m, 3H), 1.69-1.65 (m, 2H), 1.65-1.58 (m, 2H), 1.56-1.47 (m, 2H). HRMS (ESI) m/z: calcd for C47H54ClFN9O6S+ [M+H]+, 926.3585; found, 926.3581.
Referring to the method of example 1, the target compound (SIAIS262036) was prepared by using Dacomitinib derivative B and intermediate LM (SIAIS151025). (yellow solid, 7.4 mg, yield 47%)1H NMR (500 MHz, MeOD) δ 9.25 (s, 1H), 8.74 (s, 1H), 7.93 (dd, J=6.6, 2.5 Hz, 1H), 7.68-7.63 (m, 1H), 7.55-7.49 (m, 1H), 7.37 (t, J=8.9 Hz, 1H), 7.29 (s, 1H), 7.09 (dd, J=18.4, 7.4 Hz, 2H), 6.99 (d, J=8.5 Hz, 1H), 6.88 (d, J=15.1 Hz, 1H), 5.08 (dd, J=12.7, 5.5 Hz, 1H), 4.26 (s, 2H), 4.18 (s, 3H), 4.10 (s, 2H), 3.81 (d, J=11.7 Hz, 2H), 3.68 (s, 1H), 3.39-3.36 (m, 8H), 3.26 (d, J=12.3 Hz, 2H), 2.88 (m, 1H), 2.75-2.70 (m, 2H), 2.54-2.51 (s, 2H), 2.30 (s, 2H), 2.17-2.07 (m, 1H). HRMS (ESI) m/z: calcd for C43H45ClFN10O7+ [M+H]+, 867.3140; found, 867.3141.
Referring to the method of example 1, the target compound (SIAIS262037) was prepared by using Dacomitinib derivative B and intermediate LM (SIAIS151086). (yellow solid, 9.1 mg, yield 54%)1H NMR (500 MHz, MeOD) δ 9.27 (s, 1H), 8.75 (d, J=2.4 Hz, 1H), 7.96-7.90 (m, 1H), 7.68-7.61 (m, 1H), 7.56 (dd, J=10.9, 4.7 Hz, 1H), 7.38 (dd, J=10.1, 7.7 Hz, 1H), 7.32 (d, J=2.1 Hz, 1H), 7.05 (t, J=7.4 Hz, 3H), 6.88 (d, J=15.6 Hz, 1H), 5.09-5.05 (m, 1H), 4.18 (d, J=2.1 Hz, 3H), 4.09 (s, 2H), 3.79 (d, J=10.9 Hz, 2H), 3.65 (s, 2H), 3.35 (d, J=5.0 Hz, 9H), 3.24 (s, 2H), 2.85 (dd, J=13.3, 4.7 Hz, 1H), 2.73 (t, J=13.9 Hz, 2H), 2.49-2.44 (m, 4H), 2.24 (s, 2H), 2.17-2.07 (m, 1H), 1.68-1.65 (m, 4H), 1.46 (s, 4H). HRMS (ESI) m/z: calcd for C48H55ClFN10O7+ [M+H]+, 937.3922; found, 937.3920.
Referring to the method of example 1, the target compound (SIAIS262052) was prepared by using Dacomitinib derivative B and intermediate LM (SIAIS172147). (yellow solid, 8.6 mg, yield 55%)1H NMR (500 MHz, MeOD) δ 9.27 (s, 1H), 8.75 (d, J=2.4 Hz, 1H), 7.96-7.90 (m, 1H), 7.68-7.61 (m, 1H), 7.56 (dd, J=10.9, 4.7 Hz, 1H), 7.38 (dd, J=10.1, 7.7 Hz, 1H), 7.32 (d, J=2.1 Hz, 1H), 7.05 (t, J=7.4 Hz, 3H), 6.88 (d, J=15.6 Hz, 1H), 5.09-5.05 (m, 1H), 4.18 (d, J=2.1 Hz, 3H), 4.09 (s, 2H), 3.79 (d, J=10.9 Hz, 2H), 3.65 (s, 2H), 3.35 (d, J=5.0 Hz, 9H), 3.24 (s, 2H), 2.85 (dd, J=13.3, 4.7 Hz, 1H), 2.73 (t, J=13.9 Hz, 2H), 2.49-2.44 (m, 4H), 2.24 (s, 2H), 2.17-2.07 (m, 1H), 1.68-1.65 (m, 4H), 1.46 (s, 4H). HRMS (ESI) m/z: calcd for C44H48ClFN9O6+ [M+H]+, 852.3395; found, 852.3391.
Referring to the method of example 1, the target compound (SIAIS249029) was prepared by using Poziotinib derivative A and intermediate LM (SIAIS151001). (yellow solid, 9.7 mg, yield 52%)1H NMR (500 MHz, MeOD) δ 8.72 (s, 1H), 7.98 (s, 1H), 7.57-7.51 (m, 3H), 7.21 (s, 1H), 7.06 (d, J=8.6 Hz, 1H), 6.94 (d, J=7.1 Hz, 1H), 4.99 (dd, J=12.6, 5.5 Hz, 1H), 4.06 (s, 3H), 3.86-3.81 (m, 2H), 3.75-3.70 (m, 2H), 3.59-3.39 (m, 5H), 2.90-2.55 (m, 6H), 2.15-2.00 (m, 4H), 1.84-1.78 (m, 2H). HRMS (ESI) m/z: calcd for C38H37Cl2FN7O8+ [M+H]+, 808.2059; found, 808.2059.
Referring to the method of example 1, the target compound (SIAIS249030) was prepared by using Poziotinib derivative A and intermediate LM (SIAIS151004). (yellow solid, 9.5 mg, yield 48%) 1H NMR (500 MHz, MeOD) δ 8.70 (s, 1H), 7.95 (d, J=6.9 Hz, 1H), 7.59-7.52 (m, 2H), 7.47-7.42 (m, 1H), 7.22 (d, J=2.0 Hz, 1H), 7.00 (t, J=8.5 Hz, 1H), 6.91 (d, J=7.1 Hz, 1H), 5.03 (dd, J=8.3, 4.3 Hz, 1H), 4.07 (d, J=2.1 Hz, 3H), 3.99 (d, J=8.6 Hz, 1H), 3.93-3.86 (m, 1H), 3.78 (t, J=5.8 Hz, 2H), 3.71 (t, J=5.3 Hz, 2H), 3.67-3.63 (m, 4H), 3.57-3.43 (m, 5H), 2.88-2.58 (m, 6H), 2.18-2.02 (m, 4H). HRMS (ESI) m/z: calcd for C40H41Cl2FN7O9+≥[M+H]+, 852.2321; found, 852.2317.
Referring to the method of example 1, the target compound (SIAIS249031) was prepared by using Poziotinib derivative A and intermediate LM (SIAIS151005). (yellow solid, 11.0 mg, yield 53%)1H NMR (500 MHz, MeOD) δ 8.70 (s, 1H), 7.98 (d, J=3.5 Hz, 1H), 7.55 (q, J=9.0 Hz, 2H), 7.48 (t, J=7.8 Hz, 1H), 7.24 (s, 1H), 7.02 (dd, J=8.6, 3.9 Hz, 1H), 6.96 (d, J=7.1 Hz, 1H), 5.04 (dd, J=12.8, 5.3 Hz, 1H), 4.09 (d, J=6.5 Hz, 3H), 3.97-3.85 (m, 2H), 3.76 (t, J=5.4 Hz, 2H), 3.71 (t, J=5.2 Hz, 2H), 3.65 (s, 1H), 3.62-3.54 (m, 4H), 3.46 (dd, J=8.1, 5.0 Hz, 2H), 2.90-2.80 (m, 1H), 2.77-2.62 (m, 5H), 2.18-2.03 (m, 4H), 1.94 (s, 1H), 1.81 (d, J=8.9 Hz, 1H). HRMS (ESI) m/z: calcd for C42H45Cl2FN7O10+ [M+H]+, 896.2584; found, 896.2581.
Referring to the method of example 1, the target compound (SIAIS249032) was prepared by using Poziotinib derivative A and intermediate LM (SIAIS151006). (yellow solid, 11.2 mg, yield 52%)1H NMR (500 MHz, MeOD) δ 8.70 (s, 1H), 8.00 (d, J=3.8 Hz, 1H), 7.58-7.54 (m, 2H), 7.54-7.47 (m, 1H), 7.25 (s, 1H), 7.07-7.02 (m, 1H), 7.03-6.96 (m, 1H), 5.03 (dd, J=12.7, 5.4 Hz, 1H), 4.09 (s, 3H), 3.97-3.87 (m, 2H), 3.76 (t, J=6.2 Hz, 2H), 3.71 (t, J=5.0 Hz, 2H), 3.62-3.56 (m, 15H), 3.46 (t, J=4.1 Hz, 2H), 2.90-2.80 (m, 1H), 2.78-2.66 (m, 4H), 2.19-2.05 (m, 3H), 1.94 (s, 1H), 1.83 (d, J=8.8 Hz, 1H). HRMS (ESI) m/z: calcd for C44H49Cl2FN7O11+ [M+H]+, 940.2846; found, 940.2841.
Referring to the method of example 1, the target compound (SIAIS249033) was prepared by using Poziotinib derivative A and intermediate LM (SIAIS151007). (yellow solid, 11.4 mg, yield 50%)1H NMR (500 MHz, MeOD) δ 8.69 (s, 1H), 7.98 (d, J=11.8 Hz, 1H), 7.59-7.53 (m, 2H), 7.49 (dd, J=15.6, 8.2 Hz, 1H), 7.24 (d, J=4.0 Hz, 1H), 7.04-6.95 (m, 2H), 5.03 (dd, J=12.9, 5.4, Hz, 1H), 4.08 (s, 3H), 3.89 (d, J=4.1 Hz, 2H), 3.76 (t, J=6.2 Hz, 2H), 3.71 (t, J=5.1 Hz, 2H), 3.69-3.48 (m, 19H), 3.47-3.42 (m, 2H), 2.91-2.80 (m, 1H), 2.77-2.66 (m, 4H), 2.19-2.06 (m, 3H), 1.97 (d, J=19.9 Hz, 1H), 1.84 (s, 1H). HRMS (ESI) m/z: calcd for C46H53Cl2FN7O12+ [M+H]+, 984.3108; found, 984.3102.
Referring to the method of example 1, the target compound (SIAIS219177) was prepared by using Poziotinib derivative A and intermediate LM (SIAIS151025). (yellow solid, 8.6 mg, yield 50%)1H NMR (500 MHz, MeOD) δ 8.71 (s, 1H), 8.05 (s, 1H), 7.63 (t, J=8.7 Hz, 2H), 7.58-7.50 (m, 3H), 7.28 (s, 1H), 5.15 (dd, J=13.2, 4.6 Hz, 1H), 4.46 (d, J=17.3 Hz, 1H), 4.40 (d, J=17.3 Hz, 1H), 4.09 (s, 3H), 3.95-3.79 (m, 2H), 3.54 (d, J=9.3 Hz, 2H), 3.36 (s, 1H), 3.11-3.04 (m, 2H), 2.94-2.84 (m, 1H), 2.77 (dd, J=15.4, 2.2 Hz, 1H), 2.58-2.46 (m, 1H), 2.42 (t, J=7.6 Hz, 2H), 2.18 (dt, J=12.8, 6.5 Hz, 1H), 2.14-2.02 (m, 2H), 1.93-1.79 (m, 2H), 1.68 (dt, J=14.7, 7.2 Hz, 2H), 1.63-1.58 (m, 2H), 1.55-1.45 (m, 2H), 1.42-1.38 (m, 2H). HRMS (ESI) m/z: calcd for C35H31Cl2FN7O7+ [M+H]+, 750.1641; found, 750.1638.
Referring to the method of example 1, the target compound (SIAIS219179) was prepared by using Poziotinib derivative A and intermediate LM (SIAIS151019). (yellow solid, 9.2 mg, yield 51%)1H NMR (500 MHz, MeOD) δ 8.71 (s, 1H), 8.05 (s, 1H), 7.63 (t, J=8.7 Hz, 2H), 7.58-7.50 (m, 3H), 7.28 (s, 1H), 5.15 (dd, J=13.2, 4.6 Hz, 1H), 4.46 (d, J=17.3 Hz, 1H), 4.40 (d, J=17.3 Hz, 1H), 4.09 (s, 3H), 3.95-3.79 (m, 2H), 3.54 (d, J=9.3 Hz, 2H), 3.36 (s, 1H), 3.11-3.04 (m, 2H), 2.94-2.84 (m, 1H), 2.77 (dd, J=15.4, 2.2 Hz, 1H), 2.58-2.46 (m, 1H), 2.42 (t, J=7.6 Hz, 2H), 2.18 (dt, J=12.8, 6.5 Hz, 1H), 2.14-2.02 (m, 2H), 1.93-1.79 (m, 2H), 1.68 (dt, J=14.7, 7.2 Hz, 2H), 1.63-1.58 (m, 2H), 1.55-1.45 (m, 2H), 1.42-1.38 (m, 2H). HRMS (ESI) m/z: calcd for C37H35Cl2FN7O7+≥[M+H]+, 778.1954; found, 778.1950.
Referring to the method of example 1, the target compound (SIAIS219180) was prepared by using Poziotinib derivative A and intermediate LM (SIAIS151020). (yellow solid, 9.7 mg, yield 53%)1H NMR (500 MHz, MeOD) δ 8.71 (s, 1H), 8.05 (s, 1H), 7.63 (t, J=8.7 Hz, 2H), 7.58-7.50 (m, 3H), 7.28 (s, 1H), 5.15 (dd, J=13.2, 4.6 Hz, 1H), 4.46 (d, J=17.3 Hz, 1H), 4.40 (d, J=17.3 Hz, 1H), 4.09 (s, 3H), 3.95-3.79 (m, 2H), 3.54 (d, J=9.3 Hz, 2H), 3.36 (s, 1H), 3.11-3.04 (m, 2H), 2.94-2.84 (m, 1H), 2.77 (dd, J=15.4, 2.2 Hz, 1H), 2.58-2.46 (m, 1H), 2.42 (t, J=7.6 Hz, 2H), 2.18 (dt, J=12.8, 6.5 Hz, 1H), 2.14-2.02 (m, 2H), 1.93-1.79 (m, 2H), 1.68 (dt, J=14.7, 7.2 Hz, 2H), 1.63-1.58 (m, 2H), 1.55-1.45 (m, 2H), 1.42-1.38 (m, 2H). HRMS (ESI) m/z: calcd for C38H37C12FN7O7+ [M+H]+, 792.2110; found, 792.2113.
Referring to the method of example 1, the target compound (SIAIS219181) was prepared by using Poziotinib derivative A and intermediate LM (SIAIS151027). (yellow solid, 9.3 mg, yield 50%)1H NMR (500 MHz, MeOD) δ 8.71 (s, 1H), 8.05 (s, 1H), 7.63 (t, J=8.7 Hz, 2H), 7.58-7.50 (m, 3H), 7.28 (s, 1H), 5.15 (dd, J=13.2, 4.6 Hz, 1H), 4.46 (d, J=17.3 Hz, 1H), 4.40 (d, J=17.3 Hz, 1H), 4.09 (s, 3H), 3.95-3.79 (m, 2H), 3.54 (d, J=9.3 Hz, 2H), 3.36 (s, 1H), 3.11-3.04 (m, 2H), 2.94-2.84 (m, 1H), 2.77 (dd, J=15.4, 2.2 Hz, 1H), 2.58-2.46 (m, 1H), 2.42 (t, J=7.6 Hz, 2H), 2.18 (dt, J=12.8, 6.5 Hz, 1H), 2.14-2.02 (m, 2H), 1.93-1.79 (m, 2H), 1.68 (dt, J=14.7, 7.2 Hz, 2H), 1.63-1.58 (m, 2H), 1.55-1.45 (m, 2H), 1.42-1.38 (m, 2H). HRMS (ESI) m/z: calcd for C39H39Cl2FN7O7+ [M+H]+, 806.2267; found, 806.2262.
Referring to the method of example 1, the target compound (SIAIS249014) was prepared by using Poziotinib derivative A and intermediate LM (SIAIS151045). (yellow solid, 6.7 mg, yield 38%)1H NMR (500 MHz, MeOD) δ 8.69 (s, 1H), 8.06 (s, 1H), 7.87 (d, J=8.1 Hz, 1H), 7.75 (t, J=7.7 Hz, 1H), 7.67 (d, J=7.3 Hz, 1H), 7.58-7.52 (m, 2H), 7.29 (s, 1H), 5.13 (dd, J=12.7, 5.5 Hz, 1H), 4.20 (s, 2H), 4.10 (s, 3H), 4.00-3.87 (m, 2H), 3.73-3.60 (m, 2H), 3.33 (s, 1H), 2.89-2.85 (m, 1H), 2.78-2.69 (m, 2H), 2.29-2.20 (m, 1H), 2.17-1.99 (m, 3H), 1.88-1.83 (m, 1H). HRMS (ESI) m/z: calcd for C35H30Cl2FN6O7S+ [M+H]+, 767.1252; found, 767.1250.
Referring to the method of example 1, the target compound (SIAIS249015) was prepared by using Poziotinib derivative A and intermediate LM (SIAIS151138B). (yellow solid, 6.9 mg, yield 38%)1H NMR (500 MHz, MeOD) δ 8.70 (s, 1H), 8.05 (s, 1H), 7.76 (d, J=6.0 Hz, 2H), 7.66-7.61 (m, 1H), 7.58-7.52 (m, 2H), 7.28 (s, 1H), 5.12 (dd, J=9.0, 3.7 Hz, 1H), 4.09 (s, 3H), 3.94-3.87 (m, 1H), 3.82 (d, J=3.8 Hz, 1H), 3.66-3.59 (m, 1H), 3.59-3.50 (m, 1H), 3.43 (t, J=7.0 Hz, 2H), 3.33 (s, 1H), 2.91 (t, J=6.9 Hz, 2H), 2.89-2.81 (m, 1H), 2.76-2.72 (m, 2H), 2.16-2.05 (m, 3H), 1.92-1.82 (m, 2H). HRMS (ESI) m/z: calcd for C36H32Cl2FN6O7S+ [M+H]+, 781.1409; found, 781.1405.
Referring to the method of example 1, the target compound (SIAIS249016) was prepared by using Poziotinib derivative A and intermediate LM (SIAIS151139B). (yellow solid, 5.7 mg, yield 31%)1H NMR (500 MHz, MeOD) δ 8.71 (d, J=1.9 Hz, 1H), 8.03 (d, J=2.8 Hz, 1H), 7.83 (d, J=8.2 Hz, 1H), 7.75 (t, J=7.7 Hz, 1H), 7.60 (t, J=7.5 Hz, 1H), 7.56-7.52 (m, 2H), 7.29 (s, 1H), 5.11 (dd, J=8.7, 3.9 Hz, 1H), 5.08 (dd, J=8.8, 4.0 Hz, 1H), 4.10 (s, 3H), 3.94 (d, J=12.9 Hz, 1H), 3.83 (s, 1H), 3.58-3.54 (m, 2H), 3.28-3.15 (m, 2H), 2.92-2.82 (m, 1H), 2.80-2.61 (m, 4H), 2.19-2.01 (m, 5H), 1.84 (s, 2H). HRMS (ESI) m/z: calcd for C37H34Cl2FN6O7S+ [M+H]+, 795.1565; found, 795.1561.
Referring to the method of example 1, the target compound (SIAIS249017) was prepared by using Poziotinib derivative A and intermediate LM (SIAIS151140B). (yellow solid, 8.5 mg, yield 46%)1H NMR (500 MHz, MeOD) δ 8.70 (s, 1H), 8.04 (s, 1H), 7.76-7.71 (m, 2H), 7.60 (dd, J=6.0, 1.8 Hz, 1H), 7.58-7.51 (m, 2H), 7.28 (s, 1H), 5.10 (dd, J=12.6, 5.5 Hz, 1H), 4.10 (s, 3H), 3.95-3.83 (m, 2H), 3.63-3.53 (m, 2H), 3.33 (s, 1H), 3.18 (t, J=6.5 Hz, 2H), 2.92-2.79 (m, 1H), 2.78-2.64 (m, 2H), 2.53 (t, J=6.8 Hz, 2H), 2.22-2.02 (m, 4H), 1.92 (d, J=17.1 Hz, 1H), 1.85-1.82 (m, 4H). HRMS (ESI) m/z: calcd for C38H36Cl2FN6O7S+ [M+H]+, 809.1722; found, 809.1720.
Referring to the method of example 1, the target compound (SIAIS249018) was prepared by using Poziotinib derivative A and intermediate LM (SIAIS151141B). (yellow solid, 6.5 mg, yield 34%)1H NMR (500 MHz, MeOD) δ 8.70 (s, 1H), 8.03 (s, 1H), 7.75-7.69 (m, 2H), 7.59 (d, J=6.6 Hz, 1H), 7.58-7.53 (m, 2H), 7.28 (s, 1H), 5.12-5.08 (m, 1H), 4.10 (s, 3H), 3.95-3.83 (m, 2H), 3.61-3.52 (m, 2H), 3.29-3.25 (m, 1H), 3.15 (t, J=7.2 Hz, 2H), 2.87-2.82 (m, 1H), 2.77-2.69 (m, 2H), 2.48 (t, J=7.4 Hz, 2H), 2.18-2.00 (m, 4H), 1.92 (d, J=18.2 Hz, 1H), 1.83-1.79 (m, 2H), 1.71-1.67 (m, 2H), 1.62-1.57 (m, 2H). HRMS (ESI) m/z: calcd for C39H38Cl2FN6O7S+ [M+H]+, 823.1878; found, 823.1870.
Referring to the method of example 1, the target compound (SIAIS249019) was prepared by using Poziotinib derivative A and intermediate LM (SIAIS151142B). (yellow solid, 9.0 mg, yield 47%)1H NMR (500 MHz, MeOD) δ 8.70 (s, 1H), 8.03 (s, 1H), 7.75-7.68 (m, 3H), 7.58 (d, J=7.3 Hz, 1H), 7.55 (s, 1H), 7.28 (s, 1H), 5.12-5.09 (m, 1H), 4.09 (s, 3H), 3.92-3.88 (m, 2H), 3.60-3.53 (m, 2H), 3.15-3.11 (m, 3H), 2.91-2.80 (m, 2H), 2.75-2.68 (m, 2H), 2.48-2.44 (m, 2H), 2.30 (t, J=7.4 Hz, 1H), 2.18-2.03 (m, 4H), 1.91 (s, 1H), 1.79-1.75 (m, 2H), 1.68-1.63 (m, 2H), 1.59-1.55 (m, 2H), 1.47-1.43 (m, 2H). HRMS (ESI) m/z: calcd for C40H40Cl2FN6O7S+ [M+H]+, 837.2035; found, 837.2031.
Referring to the method of example 1, the target compound (SIAIS219164) was prepared by using Poziotinib derivative A and intermediate LM (SIAIS17109). (yellow solid, 8.4 mg, yield 48%)1H NMR (500 MHz, MeOD) δ 8.69 (s, 1H), 8.02 (d, J=2.3 Hz, 1H), 7.80 (dd, J=7.7, 2.1 Hz, 1H), 7.74 (d, J=7.5 Hz, 1H), 7.55 (q, J=8.9 Hz, 3H), 7.27 (s, 1H), 5.20-5.15 (m, 1H), 4.57 (dd, J=17.5, 2.7 Hz, 1H), 4.50 (dd, J=17.3, 3.7 Hz, 1H), 4.10 (s, 3H), 4.00 (t, J=9.9 Hz, 2H), 3.85-3.81 (m, 2H), 3.62-3.51 (m, 2H), 3.35 (s, 1H), 2.97-2.87 (m, 1H), 2.78 (d, J=17.5 Hz, 1H), 2.55-2.51 (m, 1H), 2.23-2.16 (m, 1H), 2.06 (s, 2H), 1.88-1.84 (m, 2H). HRMS (ESI) m/z: calcd for C35H32Cl2FN6O6S+ [M+H]+, 753.1460; found, 753.1455.
Referring to the method of example 1, the target compound (SIAIS219165) was prepared by using Poziotinib derivative A and intermediate LM (SIAIS171086). (yellow solid, 9.4 mg, yield 53%)1H NMR (500 MHz, MeOD) δ 8.70 (s, 1H), 8.01 (d, J=3.1 Hz, 1H), 7.75-7.70 (m, 1H), 7.68 (d, J=7.5 Hz, 1H), 7.58-7.53 (m, 3H), 7.27 (s, 1H), 5.17 (dd, J=13.2, 5.0 Hz, 1H), 4.49 (d, J=16.9 Hz, 1H), 4.44 (d, J=16.8 Hz, 1H), 4.09 (s, 3H), 3.88 (s, 1H), 3.78-3.75 (m, 1H), 3.57-3.53 (m, 1H), 3.46 (d, J=10.0 Hz, 1H), 3.40-3.33 (m, 3H), 2.94-2.86 (m, 1H), 2.81-2.76 (m, 3H), 2.60-2.47 (m, 1H), 2.23-2.15 (m, 1H), 2.05 (s, 2H), 1.83 (s, 2H). HRMS (ESI) m/z: calcd for C36H34Cl2FN6O6S+ [M+H]+, 767.1616; found, 767.1611.
Referring to the method of example 1, the target compound (SIAIS219166) was prepared by using Poziotinib derivative A and intermediate LM (SIAIS171089). (yellow solid, 9.7 mg, yield 54%)1H NMR (500 MHz, MeOD) δ 8.71 (d, J=2.4 Hz, 1H), 7.99 (d, J=4.3 Hz, 1H), 7.71 (d, J=8.3 Hz, 1H), 7.69-7.63 (m, 1H), 7.60-7.53 (m, 3H), 7.27 (d, J=3.4 Hz, 1H), 5.13 (dd, J=14.7, 5.1 Hz, 1H), 4.48 (d, J=12.8 Hz, 1H), 4.42 (d, J=12.8 Hz, 1H), 4.10 (d, J=1.6 Hz, 3H), 3.88 (s, 1H), 3.73 (t, J=20.2 Hz, 1H), 3.58-3.41 (m, 2H), 3.18-3.15 (m, 3H), 2.95-2.84 (m, 1H), 2.81-2.74 (m, 1H), 2.64-2.50 (m, 3H), 2.19-2.15 (m, 1H), 2.05-1.94 (m, 4H), 1.81 (s, 2H). HRMS (ESI) m/z: calcd for C37H36Cl2FN6O6S+ [M+H]+, 781.1773; found, 781.1770.
Referring to the method of example 1, the target compound (SIAIS219167) was prepared by using Poziotinib derivative A and intermediate LM (SIAIS171079). (yellow solid, 9.2 mg, yield 50%)1H NMR (500 MHz, MeOD) δ 8.71 (s, 1H), 8.01 (d, J=6.7 Hz, 1H), 7.65 (dd, J=14.9, 6.9 Hz, 2H), 7.58-7.54 (m, 3H), 7.28 (s, 1H), 5.14 (dd, J=13.4, 5.1 Hz, 1H), 4.48 (d, J=17.3 Hz, 1H), 4.41 (d, J=17.3 Hz, 1H), 4.10 (s, 3H), 3.92-3.76 (m, 2H), 3.59-3.47 (m, 2H), 3.34 (s, 1H), 3.19-3.04 (m, 2H), 2.94-2.82 (m, 1H), 2.81-2.77 (m, 1H), 2.57-2.42 (m, 3H), 2.22-2.14 (m, 1H), 2.08-1.72 (m, 8H). HRMS (ESI) m/z: calcd for C38H38Cl2FN6O6S+ [M+H]+, 795.1929; found, 795.1925.
Referring to the method of example 1, the target compound (SIAIS219168) was prepared by using Poziotinib derivative A and intermediate LM (SIAIS171091). (yellow solid, 10.1 mg, yield 54%)1H NMR (500 MHz, MeOD) δ 8.71 (s, 1H), 8.04 (s, 1H), 7.68-7.62 (m, 2H), 7.59-7.50 (m, 3H), 7.28 (s, 1H), 5.18-5.11 (m, 1H), 4.47 (d, J=17.3 Hz, 1H), 4.41 (d, J=17.4 Hz, 1H), 4.09 (s, 3H), 3.87-3.81 (m, 2H), 3.53-3.50 (m, 2H), 3.33 (s, 1H), 3.12-3.04 (m, 2H), 2.93-2.83 (m, 1H), 2.79-2.75 (m, 1H), 2.55-2.50 (m, 1H), 2.41 (t, J=7.3 Hz, 2H), 2.23-2.14 (m, 1H), 2.07 (s, 2H), 1.87-1.81 (m, 2H), 1.69-1.65 (m, 4H), 1.55-1.49 (m, 2H). HRMS (ESI) m/z: calcd for C39H40Cl2FN6O6S+ [M+H]+, 809.2086; found, 809.2084.
Referring to the method of example 1, the target compound (SIAIS219169) was prepared by using Poziotinib derivative A and intermediate LM (SIAIS171092). (yellow solid, 9.8 mg, yield 52%)1H NMR (500 MHz, MeOD) δ 8.71 (s, 1H), 8.05 (s, 1H), 7.63 (t, J=8.7 Hz, 2H), 7.58-7.50 (m, 3H), 7.28 (s, 1H), 5.15 (dd, J=13.2, 4.6 Hz, 1H), 4.46 (d, J=17.3 Hz, 1H), 4.40 (d, J=17.3 Hz, 1H), 4.09 (s, 3H), 3.95-3.79 (m, 2H), 3.54 (d, J=9.3 Hz, 2H), 3.36 (s, 1H), 3.11-3.04 (m, 2H), 2.94-2.84 (m, 1H), 2.77 (dd, J=15.4, 2.2 Hz, 1H), 2.58-2.46 (m, 1H), 2.42 (t, J=7.6 Hz, 2H), 2.18 (dt, J=12.8, 6.5 Hz, 1H), 2.14-2.02 (m, 2H), 1.93-1.79 (m, 2H), 1.68 (dt, J=14.7, 7.2 Hz, 2H), 1.63-1.58 (m, 2H), 1.55-1.45 (m, 2H), 1.42-1.38 (m, 2H). HRMS (ESI) m/z: calcd for C40H41Cl2FN6O6S+ [M+H]+, 823.2242; found, 823.2238.
Referring to the method of example 1, the target compound (SIAIS249024) was prepared by using Poziotinib derivative A and intermediate LM (SIAIS151010). (white solid, 11.2 mg, yield 48%)1H NMR (500 MHz, MeOD) δ 9.65 (s, 1H), 8.70 (s, 1H), 8.16 (d, J=18.4 Hz, 1H), 7.56-7.48 (m, 6H), 7.29 (d, J=4.3 Hz, 1H), 4.96 (s, 1H), 4.69 (s, 1H), 4.56 (dd, J=19.5, 11.0 Hz, 2H), 4.49 (s, 2H), 4.44-4.35 (m, 3H), 4.23-4.05 (m, 6H), 3.94-3.74 (m, 8H), 3.57 (d, J=7.5 Hz, 2H), 2.59-2.56 (m, 1H), 2.55 (s, 3H), 2.28-2.16 (m, 2H), 2.13-2.02 (m, 2H), 1.89-1.84 (m, 2H), 1.03 (s, 9H). HRMS (ESI) m/z: calcd for C48H56Cl2FN8O9S+ [M+H]+, 1009.3247; found, 1009.3243.
Referring to the method of example 1, the target compound (SIAIS249025) was prepared by using Poziotinib derivative A and intermediate LM (SIAIS151002). (white solid, 12.1 mg, yield 51%)1H NMR (500 MHz, MeOD) δ 9.50 (s, 1H), 8.70 (s, 1H), 8.14 (d, J=3.5 Hz, 1H), 7.55-7.46 (m, 6H), 7.30 (s, 1H), 4.96 (d, J=3.3 Hz, 1H), 4.64 (s, 1H), 4.56 (dd, J=18.3, 11.0 Hz, 3H), 4.49 (s, 1H), 4.38 (d, J=15.7 Hz, 1H), 4.09 (s, 3H), 3.92 (dd, J=33.3, 9.7 Hz, 3H), 3.78-3.73 (m, 6H), 3.65-3.50 (m, 8H), 2.73 (dd, J=13.3, 6.6 Hz, 2H), 2.61-2.49 (m, 6H), 2.24-2.18 (m, 2H), 2.15-2.03 (m, 2H), 1.97-1.77 (m, 2H), 1.03 (s, 9H). HRMS (ESI) m/z: calcd for C50H60Cl2FN8O9S+ [M+H]+, 1037.3560; found, 1037.3555.
Referring to the method of example 1, the target compound (SIAIS249026) was prepared by using Poziotinib derivative A and intermediate LM (SIAIS151003). (white solid, 12.4 mg, yield 50%)1H NMR (500 MHz, MeOD) δ 9.47 (s, 1H), 8.71 (s, 1H), 8.14 (s, 1H), 7.55-7.46 (m, 6H), 7.30 (s, 1H), 4.95 (s, 1H), 4.64 (s, 1H), 4.60-4.51 (m, 3H), 4.49 (s, 1H), 4.38 (d, J=15.7 Hz, 1H), 4.09 (s, 3H), 3.96 (d, J=3.9 Hz, 1H), 3.89 (d, J=10.9 Hz, 2H), 3.82-3.70 (m, 6H), 3.67-3.57 (m, 12H), 2.77-2.69 (m, 2H), 2.61-2.48 (m, 6H), 2.25-2.15 (m, 2H), 2.14-2.05 (m, 2H), 1.91-1.86 (m, 2H), 1.03 (s, 9H). HRMS (ESI) m/z: calcd for Cs2H64Cl2FN8O10S+ [M+H]+, 1081.3822; found, 1081.3820.
Referring to the method of example 1, the target compound (SIAIS249027) was prepared by using Poziotinib derivative A and intermediate LM (SIAIS151008). (white solid, 12.1 mg, yield 47%)1H NMR (500 MHz, MeOD) δ 9.53 (s, 1H), 8.71 (s, 1H), 8.15 (s, 1H), 7.55-7.46 (m, 6H), 7.31 (s, 1H), 4.99-4.93 (m, 1H), 4.64 (s, 1H), 4.60-4.52 (m, 2H), 4.49 (s, 1H), 4.38 (d, J=15.7 Hz, 1H), 4.09 (s, 3H), 3.98 (s, 1H), 3.88 (d, J=10.7 Hz, 2H), 3.82-3.70 (m, 6H), 3.62 (d, J=6.3 Hz, 16H), 2.78-2.65 (m, 2H), 2.62-2.53 (m, 5H), 2.51-2.45 (m, 1H), 2.28-2.15 (m, 2H), 2.15-2.05 (m, 2H), 1.98-1.78 (m, 2H), 1.03 (s, 9H). HRMS (ESI) m/z: calcd for C54H68Cl2FN8O1S+ [M+H]+, 1125.4084; found, 1125.4081.
Referring to the method of example 1, the target compound (SIAIS249028) was prepared by using Poziotinib derivative A and intermediate LM (SIAIS151009). (white solid, 13.1 mg, yield 49%)1H NMR (500 MHz, MeOD) δ 9.23 (s, 1H), 8.71 (s, 1H), 8.16 (s, 1H), 7.57-7.43 (m, 6H), 7.31 (s, 1H), 5.00-4.94 (m, 1H), 4.64 (s, 1H), 4.61-4.47 (m, 4H), 4.37 (d, J=15.6 Hz, 1H), 4.10 (d, J=9.6 Hz, 3H), 4.01-3.95 (m, 1H), 3.88 (d, J=11.0 Hz, 2H), 3.83-3.70 (m, 6H), 3.66-3.56 (m, 22H), 2.75-2.69 (m, 2H), 2.61-2.54 (m, 1H), 2.52-2.45 (m, 5H), 2.24-2.16 (m, 2H), 2.09-2.04 (m, 2H), 1.96-1.90 (m, 1H), 1.84-1.79 (m, 1H). HRMS (ESI) m/z: calcd for C56H72Cl2FN8O12S+ [M+H]+, 1169.4346; found, 1169.4341.
Referring to the method of example 1, the target compound (SIAIS249020) was prepared by using Poziotinib derivative A and intermediate LM (SIAIS074011). (white solid, 7.7 mg, yield 35%)1H NMR (500 MHz, MeOD) δ 9.24 (d, J=3.2 Hz, 1H), 8.71 (d, J=2.6 Hz, 1H), 8.05 (s, 1H), 7.58-7.54 (m, 2H), 7.50 (d, J=8.2 Hz, 2H), 7.45 (d, J=8.2 Hz, 2H), 7.28 (s, 1H), 4.91 (s, 1H), 4.61 (d, J=2.0 Hz, 1H), 4.56 (dd, J=18.6, 10.6 Hz, 2H), 4.49 (s, 1H), 4.38 (d, J=15.5 Hz, 1H), 4.09 (t, J=5.2 Hz, 3H), 3.89 (d, J=10.2 Hz, 3H), 3.80 (dd, J=11.0, 3.9 Hz, 1H), 3.59 (d, J=9.8 Hz, 2H), 2.79-2.70 (m, 2H), 2.69-2.61 (m, 1H), 2.57 (dd, J=14.1, 7.4 Hz, 1H), 2.52 (d, J=8.1 Hz, 3H), 2.26-2.14 (m, 2H), 2.12-2.05 (m, 2H), 1.94 (s, 1H), 1.84 (s, 1H), 1.04 (s, 9H). HRMS (ESI) m/z: calcd for C46H52Cl2FN8O7S+ [M+H]+, 949.3035; found, 949.3032.
Referring to the method of example 1, the target compound (SIAIS249021) was prepared by using Poziotinib derivative A and intermediate LM (SIAIS074013). (white solid, 8.1 mg, yield 36%)1H NMR (500 MHz, MeOD) δ 9.32 (s, 1H), 8.71 (s, 1H), 8.05 (s, 1H), 7.58-7.53 (m, 2H), 7.50 (d, J=8.4 Hz, 2H), 7.46 (dd, J=10.9, 5.8 Hz, 2H), 7.28 (s, 1H), 4.91 (s, 1H), 4.63 (d, J=3.9 Hz, 1H), 4.60-4.52 (m, 2H), 4.49 (s, 1H), 4.37 (d, J=15.6 Hz, 1H), 4.09 (s, 3H), 3.90 (d, J=11.1 Hz, 3H), 3.80 (dd, J=10.9, 3.9 Hz, 1H), 3.62-3.53 (m, 2H), 2.53 (d, J=9.0 Hz, 3H), 2.48 (d, J=6.7 Hz, 2H), 2.38-2.32 (m, 2H), 2.23-2.18 (m, 2H), 2.09-2.06 (m, 2H), 1.90 (d, J=51.0 Hz, 2H), 1.69-1.64 (m, 4H), 1.04 (s, 9H). HRMS (ESI) m/z: calcd for C48H56Cl2FN8O7S+ [M+H]+, 977.3348; found, 977.3343.
Referring to the method of example 1, the target compound (SIAIS249022) was prepared by using Poziotinib derivative A and intermediate LM (SIAIS074015). (white solid, 9.7 mg, yield 42%)1H NMR (500 MHz, MeOD) δ 9.36 (s, 1H), 8.71 (s, 1H), 8.07 (s, 1H), 7.58-7.53 (m, 2H), 7.53-7.49 (m, 2H), 7.46 (t, J=6.5 Hz, 2H), 7.28 (s, 1H), 4.92 (s, 1H), 4.63 (s, 1H), 4.60-4.47 (m, 3H), 4.38 (d, J=15.6 Hz, 1H), 4.09 (s, 3H), 3.90 (q, J=17.7 Hz, 3H), 3.80 (dd, J=11.0, 3.9 Hz, 1H), 3.57 (dd, J=11.1, 6.7 Hz, 2H), 2.52 (s, 3H), 2.47-2.42 (m, 2H), 2.34-2.14 (m, 4H), 2.12-2.05 (m, 2H), 1.96-1.80 (m, 2H), 1.69-1.60 (m, 4H), 1.43-1.35 (m, 4H), 1.03 (s, 9H). HRMS (ESI) m/z: calcd for C50H60Cl2FN8O7S+ [M+H]+, 1005.3661; found, 1005.3656.
Referring to the method of example 1, the target compound (SIAIS249023) was prepared by using Poziotinib derivative A and intermediate LM (SIAIS074019). (white solid, 10.8 mg, yield 45%)1H NMR (500 MHz, MeOD) δ 9.31 (s, 1H), 8.71 (s, 1H), 8.08 (d, J=3.8 Hz, 1H), 7.59-7.54 (m, 2H), 7.51 (q, J=5.6 Hz, 2H), 7.46 (t, J=7.0 Hz, 2H), 7.29 (s, 1H), 4.92 (dd, J=6.9, 3.5 Hz, 1H), 4.64 (s, 1H), 4.61-4.48 (m, 3H), 4.42-4.35 (m, 1H), 4.10 (s, 3H), 3.95-3.84 (m, 3H), 3.80 (dd, J=10.9, 3.8 Hz, 1H), 3.62-3.52 (m, 2H), 2.56-2.52 (m, 3H), 2.49-2.41 (m, 2H), 2.33-2.15 (m, 4H), 2.11-2.07 (m, 2H), 1.94-1.77 (m, 2H), 1.61 (d, J=6.2 Hz, 4H), 1.35 (s, 8H), 1.03 (s, 9H). HRMS (ESI) m/z: calcd for Cs2H64Cl2FN8O7S+ [M+H]+, 1033.3974; found, 1033.3971.
Referring to the method of example 1, the target compound (SIAIS184164) was prepared by using Gefitinib derivative A and intermediate LM (SIAIS151045). (yellow solid, 6.9 mg, yield 40%) 1H NMR (500 MHz, MeOD) δ 8.74 (s, 1H), 8.08 (d, J=3.5 Hz, 1H), 8.00-7.96 (m, 1H), 7.96 (s, 1H), 7.80-7.75 (m, 1H), 7.73 (s, 1H), 7.72-7.67 (m, 1H), 7.38 (t, J=8.9 Hz, 1H), 7.27 (s, 1H), 5.14 (dd, J=12.6, 5.6 Hz, 1H), 4.42 (d, J=5.6 Hz, 2H), 4.10 (t, J=3.4 Hz, 3H), 3.74 (s, 2H), 3.50 (s, 2H), 3.29-2.95 (m, 8H), 2.89-2.82 (m, 1H), 2.76-2.71 (m, 2H), 2.45 (s, 2H), 2.15 (s, 1H). HRMS (ESI) m/z: calcd for C37H36ClFN7O7S+ [M+H]+, 776.2064; found, 776.2061.
Referring to the method of example 1, the target compound (SIAIS184165) was prepared by using Gefitinib derivative A and intermediate LM (SIAIS151139B). (yellow solid, 7.8 mg, yield 43%) 1H NMR (500 MHz, MeOD) δ 8.73 (s, 1H), 8.09 (s, 1H), 7.98 (dd, J=6.6, 2.6 Hz, 1H), 7.82 (d, J=8.0 Hz, 1H), 7.77-7.74 (m, 1H), 7.70 (ddd, J=9.0, 4.1, 2.6 Hz, 1H), 7.62 (d, J=7.0 Hz, 1H), 7.37 (t, J=8.9 Hz, 1H), 7.26 (s, 1H), 5.11 (dd, J=12.6, 5.4 Hz, 1H), 4.42 (t, J=5.6 Hz, 2H), 4.09 (s, 3H), 3.82-3.58 (m, 2H), 3.49 (t, J=7.5 Hz, 2H), 3.30-3.20 (m, 8H), 2.88-2.82 (m, 1H), 2.75-2.64 (m, 4H), 2.49-2.41 (m, 2H), 2.17-2.06 (m, 3H). HRMS (ESI) m/z: calcd for C39H40ClFN7O7S+ [M+H]+, 804.2377; found, 804.2374.
Referring to the method of example 1, the target compound (SIAIS184166) was prepared by using Gefitinib derivative A and intermediate LM (SIAIS151141B). (yellow solid, 9.5 mg, yield 51%) 1H NMR (500 MHz, MeOD) δ 8.74 (s, 1H), 8.11 (s, 1H), 7.98 (dd, J=6.6, 2.6 Hz, 1H), 7.75-7.68 (m, 3H), 7.59 (dd, J=6.7, 1.3 Hz, 1H), 7.37 (t, J=8.9 Hz, 1H), 7.26 (s, 1H), 5.11 (dd, J=12.7, 5.5 Hz, 1H), 4.43 (t, J=5.5 Hz, 2H), 4.09 (s, 3H), 3.73 (d, J=40.0 Hz, 2H), 3.50 (t, J=7.3 Hz, 2H), 3.31-3.01 (m, 8H), 2.89-2.83 (m, 1H), 2.78-2.67 (m, 2H), 2.48-2.43 (m, 4H), 2.18-2.09 (m, 1H), 1.86-1.78 (m, 2H), 1.77-1.67 (m, 2H), 1.61-1.58 (m, 2H). HRMS (ESI) m/z: calcd for C41H44ClFN7O7S+ [M+H]+, 832.2690; found, 832.2687.
Referring to the method of example 1, the target compound (SIAIS184168) was prepared by using Gefitinib derivative A and intermediate LM (SIAIS1204137). (yellow solid, 8.2 mg, yield 45%) 1H NMR (500 MHz, MeOD) δ 8.73 (s, 1H), 8.05 (s, 1H), 8.02-7.97 (m, 1H), 7.71 (dd, J=10.3, 3.7 Hz, 3H), 7.57 (s, 1H), 7.39-7.35 (m, 1H), 7.22 (s, 1H), 5.08 (dd, J=12.6, 5.4 Hz, 1H), 4.45-4.41 (m, 4H), 4.08 (s, 3H), 3.93-3.90 (m, 2H), 3.56-3.53 (m, 4H), 3.38-3.34 (m, 8H), 2.84-2.75 (m, 1H), 2.69-2.65 (m, 2H), 2.46 (s, 2H), 2.11 (d, J=5.2 Hz, 1H). HRMS (ESI) m/z: calcd for C39H40ClFN7O8S+ [M+H]+, 820.2326; found, 820.2321.
Referring to the method of example 1, the target compound (SIAIS184169) was prepared by using Gefitinib derivative A and intermediate LM (SIAIS1204141). (yellow solid, 9.1 mg, yield 45%) 1H NMR (500 MHz, MeOD) δ 8.71 (s, 1H), 8.01-7.95 (m, 2H), 7.73-7.67 (m, 2H), 7.64 (s, 1H), 7.48 (dd, J=6.0, 2.0 Hz, 1H), 7.37 (dd, J=8.9, 3.4 Hz, 1H), 7.20 (s, 1H), 5.12-5.08 (m, 1H), 4.41-4.37 (m, 2H), 4.37-4.25 (m, 2H), 4.07 (s, 3H), 3.84 (t, J=6.0 Hz, 2H), 3.72-3.61 (m, 14H), 3.56-3.47 (m, 4H), 3.05 (s, 1H), 2.89-2.83 (m, 1H), 2.78-2.60 (m, 3H), 2.45 (s, 2H), 2.15-2.08 (m, 1H). HRMS (ESI) m/z: calcd for C43H48ClFN7O10S+ [M+H]+, 908.2850; found, 908.2846.
Referring to the method of example 1, the target compound (SIAIS184170) was prepared by using Gefitinib derivative A and intermediate LM (SIAIS1204149). (yellow solid, 9.2 mg, yield 41%) 1H NMR (500 MHz, MeOD) δ 8.72 (s, 1H), 8.06 (s, 1H), 7.98 (dt, J=6.0, 3.0 Hz, 1H), 7.73-7.70 (m, 1H), 7.68 (dd, J=7.1, 4.7 Hz, 2H), 7.56-7.51 (m, 1H), 7.39-7.33 (m, 1H), 7.23 (s, 1H), 5.10 (dd, J=12.7, 5.5 Hz, 1H), 4.41 (dd, J=11.5, 5.9 Hz, 2H), 4.39-4.27 (m, 2H), 4.08 (s, 3H), 3.80 (t, J=6.1 Hz, 2H), 3.69-3.60 (m, 18H), 3.57-3.41 (m, 4H), 3.37-3.32 (m, 2H), 3.29 (s, 2H), 3.26-3.04 (m, 2H), 2.89-2.83 (m, 1H), 2.79-2.63 (m, 2H), 2.50-2.42 (m, 2H), 2.17-2.09 (m, 1H). HRMS (ESI) m/z: calcd for C47H56ClFN7O12S+ [M+H]+, 996.3375; found, 996.3371.
Referring to the method of example 1, the target compound (SIAIS184184) was prepared by using Gefitinib derivative A and intermediate LM (SIAIS171090). (yellow solid, 6.7 mg, yield 39%) 1H NMR (500 MHz, MeOD) δ 8.44 (s, 1H), 8.00 (dd, J=6.7, 2.5 Hz, 1H), 7.78 (d, J=7.8 Hz, 1H), 7.75-7.73 (m, 2H), 7.69-7.64 (m, 1H), 7.55 (t, J=7.6 Hz, 1H), 7.26 (t, J=8.9 Hz, 1H), 7.18 (s, 1H), 5.17 (dd, J=13.3, 5.1 Hz, 1H), 4.55 (d, J=17.6 Hz, 1H), 4.48 (d, J=17.3 Hz, 1H), 4.25 (d, J=6.0 Hz, 2H), 4.00 (s, 3H), 3.57 (d, J=5.4 Hz, 2H), 3.53 (d, J=3.7 Hz, 2H), 3.35-3.30 (m, 8H), 2.92-2.88 (m, 1H), 2.81-2.76 (m, 1H), 2.58-2.51 (m, 1H), 2.21-2.17 (m, 1H), 2.08 (d, J=7.2 Hz, 2H). HRMS (ESI) m/z: calcd for C37H38ClFN7O6S+ [M+H]+, 762.2271; found, 762.2268.
Referring to the method of example 1, the target compound (SIAIS184185) was prepared by using Gefitinib derivative A and intermediate LM (SIAIS171089). (yellow solid, 7.2 mg, yield 41%) 1H NMR (500 MHz, MeOD) δ 8.46 (s, 1H), 8.02-7.98 (m, 1H), 7.75 (s, 1H), 7.68 (dd, J=19.3, 7.6 Hz, 3H), 7.54 (t, J=7.7 Hz, 1H), 7.27 (t, J=8.9 Hz, 1H), 7.19 (s, 1H), 5.18-5.14 (m, 1H), 4.48 (d, J=17.3 Hz, 1H), 4.42 (d, J=17.2 Hz, 1H), 4.25 (t, J=5.8 Hz, 2H), 4.01 (s, 3H), 3.69-3.65 (m, 2H), 3.60-3.36 (m, 8H), 3.15-3.10 (m, 2H), 2.89-2.84 (m, 1H), 2.79-2.75 (m, 1H), 2.65 (s, 2H), 2.55 (d, J=7.0 Hz, 1H), 2.19 (s, 1H), 2.11 (s, 2H), 1.95 (t, J=7.1 Hz, 2H). HRMS (ESI) m/z: calcd for C39H42ClFN7O6S+ [M+H]+, 790.2584; found, 790.2581.
Referring to the method of example 1, the target compound (SIAIS184186) was prepared by using Gefitinib derivative A and intermediate LM (SIAIS171091). (yellow solid, 8.2 mg, yield 45%) 1H NMR (500 MHz, MeOD) δ 8.45 (s, 1H), 8.00 (d, J=6.7 Hz, 1H), 7.74 (s, 1H), 7.65 (dd, J=19.0, 8.2 Hz, 3H), 7.52 (t, J=7.7 Hz, 1H), 7.26 (t, J=8.9 Hz, 1H), 7.18 (s, 1H), 5.17-5.13 (m, 1H), 4.46 (d, J=17.6 Hz, 1H), 4.40 (d, J=17.2 Hz, 1H), 4.24 (t, J=5.9 Hz, 2H), 4.00 (s, 3H), 3.79-3.65 (m, 2H), 3.63-3.33 (m, 8H), 3.07 (d, J=3.5 Hz, 2H), 2.92-2.86 (m, 1H), 2.79-2.75 (m, 1H), 2.60 (t, J=7.3 Hz, 2H), 2.38 (d, J=7.2 Hz, 1H), 2.22-2.08 (m, 3H), 1.71-1.67 (m, 2H), 1.65-1.59 (m, 2H), 1.58-1.49 (m, 2H). HRMS (ESI) m/z: calcd for C41H46ClFN7O6S+ [M+H]+, 818.2897; found, 818.2893.
Referring to the method of example 1, the target compound (SIAIS262085) was prepared by using Gefitinib derivative B and intermediate LM (SIAIS171090). (yellow solid, 10.1 mg, yield 43%) 1H NMR (500 MHz, MeOD) δ 8.74 (s, 1H), 8.12 (s, 1H), 7.98 (dd, J=6.6, 2.6 Hz, 1H), 7.83 (d, J=7.7 Hz, 1H), 7.78 (d, J=7.5 Hz, 1H), 7.75-7.68 (m, 1H), 7.58 (t, J=7.7 Hz, 1H), 7.37 (t, J=8.9 Hz, 1H), 7.26 (s, 1H), 5.18 (dd, J=13.4, 5.1 Hz, 1H), 4.57 (d, J=17.5 Hz, 1H), 4.50 (dd, J=14.7, 4.1 Hz, 1H), 4.43 (t, J=5.5 Hz, 2H), 4.11 (s, 3H), 3.99-3.91 (m, 5H), 3.58-3.52 (m, 4H), 3.30-3.13 (m, 8H), 2.94-2.87 (m, 1H), 2.81-2.77 (m, 1H), 2.61-2.51 (m, 1H), 2.45 (s, 3H), 2.29-2.14 (m, 4H). HRMS (ESI) m/z: calcd for C42H47ClFN8O6S+ [M+H]+, 845.3006; found, 845.3001.
Referring to the method of example 1, the target compound (SIAIS262086) was prepared by using Gefitinib derivative B and intermediate LM (SIAIS171079). (yellow solid, 11.2 mg, yield 45%) 1H NMR (500 MHz, MeOD) δ 8.74 (s, 1H), 8.11 (s, 1H), 7.98 (dd, J=6.6, 2.6 Hz, 1H), 7.75-7.68 (m, 1H), 7.66 (t, J=7.0 Hz, 2H), 7.54 (t, J=7.7 Hz, 1H), 7.37 (t, J=8.9 Hz, 1H), 7.26 (s, 1H), 5.17 (dd, J=13.3, 5.1 Hz, 1H), 4.48 (d, J=17.3 Hz, 1H), 4.46-4.39 (m, 3H), 4.11 (s, 3H), 3.92 (d, J=12.6 Hz, 2H), 3.60 (s, 2H), 3.45-3.41 (m, 5H), 3.18-3.11 (m, 8H), 2.94-2.89 (m, 1H), 2.81-2.76 (m, 1H), 2.57-2.43 (m, 7H), 2.24-2.18 (m, 3H), 1.79-1.71 (m, 4H). HRMS (ESI) m/z: calcd for C45H53ClFN8O6S+ [M+H]+, 887.3476; found, 887.3471.
Referring to the method of example 1, the target compound (SIAIS262087) was prepared by using Gefitinib derivative B and intermediate LM (SIAIS171091). (yellow solid, 11.3 mg, yield 45%) 1H NMR (500 MHz, MeOD) δ 8.74 (s, 1H), 8.02 (s, 1H), 7.96-7.93 (m, 1H), 7.69-7.64 (m, 3H), 7.54 (t, J=7.7 Hz, 1H), 7.37 (t, J=8.9 Hz, 1H), 7.27 (s, 1H), 5.17 (dd, J=13.3, 5.2 Hz, 1H), 4.48 (d, J=17.4 Hz, 1H), 4.42 (d, J=14.3 Hz, 1H), 4.39 (t, J=3.9 Hz, 2H), 4.10 (s, 3H), 3.85 (d, J=49.3 Hz, 6H), 3.45 (d, J=7.2 Hz, 2H), 3.30-3.12 (m, 6H), 3.11-3.02 (m, 2H), 2.96-2.86 (m, 1H), 2.82-2.77 (m, 1H), 2.59-2.50 (m, 1H), 2.49-2.36 (m, 6H), 2.22-2.13 (m, 3H), 1.71-1.58 (m, 4H), 1.55-1.51 (m, 2H). HRMS (ESI) m/z: calcd for C46H55ClFN8O6S+ [M+H]+, 901.3632; found, 901.3632.
Referring to the method of example 1, the target compound (SIAIS184093) was prepared by using Afatinib derivative A and intermediate LM (SIAIS151045). (white solid, 12.2 mg, yield 43%) 1H NMR (500 MHz, MeOD) δ 9.16 (s, 1H), 8.72 (s, 1H), 7.90 (dd, J=6.6, 2.6 Hz, 1H), 7.80 (d, J=7.9 Hz, 1H), 7.75 (t, J=7.6 Hz, 1H), 7.68 (d, J=7.2 Hz, 1H), 7.66-7.60 (m, 1H), 7.36 (t, J=8.9 Hz, 1H), 7.21 (s, 1H), 5.29 (s, 1H), 5.16 (dd, J=12.5, 5.5 Hz, 1H), 4.26 (s, 2H), 4.02 (qd, J=10.7, 4.1 Hz, 2H), 3.89-3.82 (m, 2H), 2.92-2.85 (m, 1H), 2.79-2.70 (m, 2H), 2.37 (dd, J=14.0, 8.0 Hz, 1H), 2.14 (dd, J=13.1, 6.1 Hz, 2H). HRMS (ESI) m/z: calcd for C33H27ClFN6O7S+ [M+H]+, 705.1329; found, 705.1326.
Referring to the method of example 1, the target compound (SIAIS184094) was prepared by using Afatinib derivative A and intermediate LM (SIAIS151139B). (white solid, 13.6 mg, yield 46%) 1H NMR (500 MHz, MeOD) δ 9.07 (s, 1H), 8.73 (s, 1H), 7.94 (dd, J=6.6, 2.4 Hz, 1H), 7.79 (d, J=8.3 Hz, 1H), 7.74 (t, J=7.6 Hz, 1H), 7.68-7.63 (m, 1H), 7.62 (d, J=7.0 Hz, 1H), 7.37 (t, J=8.9 Hz, 1H), 7.22 (s, 1H), 5.32 (s, 1H), 5.08 (dd, J=12.7, 5.3 Hz, 1H), 4.18 (d, J=10.3 Hz, 1H), 4.04 (dd, J=13.3, 7.3 Hz, 2H), 3.91 (dt, J=13.2, 6.6 Hz, 1H), 2.85-2.78 (m, 3H), 2.76-2.66 (m, 2H), 2.48-242 (m, 1H), 2.29 (s, 1H), 2.25-2.17 (m, 2H), 2.10 (s, 1H). HRMS (ESI) m/z: calcd for C35H31ClFN6O7S+ [M+H]+, 733.1642; found, 733.1640.
Referring to the method of example 1, the target compound (SIAIS184095) was prepared by using Afatinib derivative A and intermediate LM (SIAIS151141B). (white solid, 14.1 mg, yield 46%) 1H NMR (500 MHz, MeOD) δ 9.05 (s, 1H), 8.72 (s, 1H), 7.93 (dd, J=6.6, 2.6 Hz, 1H), 7.71-7.67 (m, 2H), 7.67-7.64 (m, 1H), 7.56-7.52 (m, 1H), 7.37 (t, J=8.9 Hz, 1H), 7.23 (s, 1H), 5.33 (d, J=4.6 Hz, 1H), 5.09 (dd, J=12.8, 5.4 Hz, 1H), 4.19 (d, J=10.6 Hz, 1H), 4.11-4.02 (m, 2H), 3.94-3.88 (m, 1H), 3.16 (t, J=7.0 Hz, 2H), 2.87-2.83 (m, 1H), 2.77-2.66 (m, 2H), 2.64-2.59 (m, 2H), 2.47-2.43 (m, 1H), 2.33-2.24 (m, 1H), 2.15-2.08 (m, 1H), 1.87-1.82 (m, 4H), 1.69-1.64 (m, 2H). HRMS (ESI) m/z: calcd for C37H35ClFN6O7S+ [M+H]+, 761.1955; found, 761.1952.
Referring to the method of example 1, the target compound (SIAIS184152) was prepared by using Afatinib derivative A and intermediate LM (SIAIS1204137). (white solid, 8.8 mg, yield 44%) 1H NMR (500 MHz, CDCl3) δ 9.34 (d, J=5.8 Hz, 2H), 9.08 (s, 1H), 8.73 (d, J=24.1 Hz, 1H), 8.64 (s, 1H), 7.97 (s, 1H), 7.86 (s, 1H), 7.55 (s, 3H), 7.42 (d, J=10.7 Hz, 1H), 7.21 (t, J=8.4 Hz, 1H), 5.31 (s, 1H), 4.97 (s, 1H), 4.18-4.12 (m, 2H), 4.06 (s, 2H), 3.93-3.74 (m, 2H), 3.29 (s, 2H), 2.85-2.78 (m, 3H), 2.63-2.57 (m, 1H), 2.21-2.15 (m, 3H), 1.47-1.42 (m, 1H). HRMS (ESI) m/z: calcd for C35H31ClFN6O8S+ [M+H]+, 749.1591; found, 749.1588.
Referring to the method of example 1, the target compound (SIAIS184153) was prepared by using Afatinib derivative A and intermediate LM (SIAIS1204139). (white solid, 10.5 mg, yield 50%) 1H NMR (500 MHz, CDCl3) δ 9.33 (d, J=5.8 Hz, 2H), 9.07 (s, 1H), 8.71 (d, J=24.1 Hz, 1H), 8.62 (s, 1H), 7.96 (s, 1H), 7.85 (s, 1H), 7.53 (s, 3H), 7.40 (d, J=10.7 Hz, 1H), 7.20 (t, J=8.4 Hz, 1H), 5.29 (s, 1H), 4.96 (s, 1H), 4.14 (d, J=15.7 Hz, 2H), 4.04 (s, 2H), 3.90-3.72 (m, 6H), 3.28 (s, 2H), 2.83-2.78 (m, 3H), 2.61-2.55 (m, 1H), 2.19-2.14 (m, 3H), 1.45-1.40 (m, 1H). HRMS (ESI) m/z: calcd for C37H35ClFN6O9S+ [M+H]+, 793.1853; found, 793.1851.
Referring to the method of example 1, the target compound (SIAIS184154) was prepared by using Afatinib derivative A and intermediate LM (SIAIS1204141). (white solid, 9.9 mg, yield 44%) 1H NMR (500 MHz, CDCl3) δ 9.78 (s, 1H), 9.41 (d, J=2.7 Hz, 1H), 9.20 (s, 1H), 9.09 (s, 1H), 8.57 (s, 1H), 8.02 (s, 1H), 7.86-7.77 (m, 1H), 7.61-7.49 (m, 2H), 7.46-7.43 (m, 2H), 7.17 (dd, J=14.7, 8.5 Hz, 1H), 5.25 (s, 1H), 5.05-4.93 (m, 1H), 4.23 (s, 2H), 4.09 (dd, J=31.1, 9.7 Hz, 2H), 3.94-3.74 (m, 8H), 3.69 (d, J=8.8 Hz, 4H), 3.21 (s, 2H), 2.88-2.83 (m, 1H), 2.83-2.72 (m, 2H), 2.54 (s, 1H), 2.17-2.12 (m, 2H). HRMS (ESI) m/z: calcd for C39H39ClFN6O10S+ [M+H]+, 837.2115; found, 837.2112.
Referring to the method of example 1, the target compound (SIAIS184155) was prepared by using Afatinib derivative A and intermediate LM (SIAIS1204147). (white solid, 9.8 mg, yield 42%) 1H NMR (500 MHz, CDCl3) δ 9.66 (s, 1H), 9.41 (s, 1H), 9.23 (s, 1H), 9.09 (s, 1H), 8.59 (s, 1H), 8.05 (s, 1H), 7.83 (s, 1H), 7.57 (d, J=4.7 Hz, 2H), 7.53-7.46 (m, 2H), 7.18 (td, J=8.6, 3.9 Hz, 1H), 5.25 (s, 1H), 5.04-4.93 (m, 1H), 4.24 (s, 2H), 4.09 (dd, J=24.2, 12.7 Hz, 2H), 3.95-3.76 (m, 8H), 3.71-3.64 (m, 8H), 3.24 (s, 2H), 2.88-2.84 (m, 1H), 2.83-2.71 (m, 2H), 2.53 (s, 1H), 2.19-2.14 (m, 2H). HRMS (ESI) m/z: calcd for C41H43ClFN6O11S+ [M+H]+, 881.2378; found, 881.2375.
Referring to the method of example 1, the target compound (SIAIS184156) was prepared by using Afatinib derivative A and intermediate LM (SIAIS1204149). (white solid, 10.2 mg, yield 41%) 1H NMR (500 MHz, CDCl3) δ 9.66 (s, 1H), 9.41 (s, 1H), 9.33 (s, 1H), 9.13 (s, 1H), 8.59 (s, 1H), 8.02 (s, 1H), 7.82 (d, J=5.2 Hz, 1H), 7.66-7.49 (m, 4H), 7.15 (t, J=8.6 Hz, 1H), 5.26 (s, 1H), 5.05-4.93 (m, 1H), 4.24 (s, 2H), 4.09-4.03 (m, 2H), 3.91-3.72 (m, 8H), 3.70-3.59 (m, 12H), 3.24 (d, J=5.7 Hz, 2H), 2.90-2.78 (m, 2H), 2.54 (s, 1H), 2.18-2.13 (m, 2H). HRMS (ESI) m/z: calcd for C43H47ClFN6O12S+ [M+H]+, 925.2640; found, 925.2636.
Referring to the method of example 1, the target compound (SIAIS1210085) was prepared by using Afatinib derivative A and intermediate LM (SIAIS171086). (yellow solid, 9.9 mg, yield 53%) 1H NMR (500 MHz, MeOD) δ 9.02 (s, 1H), 8.70 (s, 1H), 7.91 (dd, J=6.6, 2.5 Hz, 1H), 7.67 (t, J=6.7 Hz, 1H), 7.63-7.60 (m, 2H), 7.54-7.50 (m, 1H), 7.33 (dd, J=11.1, 6.7 Hz, 1H), 7.23 (s, 1H), 5.35-5.30 (m, 1H), 5.17-5.13 (m, 1H), 4.45 (d, J=17.2 Hz, 1H), 4.41 (d, J=17.2 Hz, 1H), 4.14 (dd, J=10.5, 4.0 Hz, 1H), 4.05-4.01 (m, 2H), 3.91-3.88 (m, 1H), 3.18 (t, J=7.2 Hz, 2H), 2.95-2.83 (m, 2H), 2.77-2.74 (m, 2H), 2.51-2.46 (m, 2H), 2.29-2.21 (m, 1H), 2.17-2.12 (m, 1H). HRMS (ESI) m/z: calcd for C34H31ClFN6O6S+ [M+H]+, 705.1693; found, 705.1691.
Referring to the method of example 1, the target compound (SIAIS1210087) was prepared by using Afatinib derivative A and intermediate LM (SIAIS171089). (yellow solid, 10.3 mg, yield 54%) 1H NMR (500 MHz, MeOD) δ 9.04 (s, 1H), 8.71 (s, 1H), 7.93 (dd, J=6.6, 2.5 Hz, 1H), 7.69 (t, J=6.7 Hz, 1H), 7.65-7.62 (m, 2H), 7.56-7.50 (m, 1H), 7.35 (dd, J=11.1, 6.7 Hz, 1H), 7.24 (s, 1H), 5.36-5.31 (m, 1H), 5.18-5.14 (m, 1H), 4.46 (d, J=17.2 Hz, 1H), 4.42 (d, J=17.2 Hz, 1H), 4.16 (dd, J=10.5, 4.0 Hz, 1H), 4.06-4.01 (m, 2H), 3.93-3.90 (m, 1H), 3.20 (t, J=7.2 Hz, 2H), 2.96-2.84 (m, 2H), 2.78-2.75 (m, 2H), 2.52-2.47 (m, 2H), 2.30-2.22 (m, 1H), 2.19-2.14 (m, 1H), 2.13-2.07 (m, 2H). HRMS (ESI) m/z: calcd for C35H33ClFN6O6S+ [M+H]+, 719.1849; found, 719.1846.
Referring to the method of example 1, the target compound (SIAIS1210089) was prepared by using Afatinib derivative A and intermediate LM (SIAIS171091). (yellow solid, 11.1 mg, yield 56%)1H NMR (500 MHz, MeOD) δ 8.95 (d, J=8.6 Hz, 1H), 8.68 (s, 1H), 7.95 (dd, J=6.6, 2.6 Hz, 1H), 7.67-7.62 (m, 2H), 7.58 (dd, J=6.8, 5.4 Hz, 1H), 7.48 (dd, J=10.0, 4.5 Hz, 1H), 7.34 (dd, J=9.2, 8.6 Hz, 1H), 7.22 (d, J=1.0 Hz, 1H), 5.31 (d, J=4.6 Hz, 1H), 5.16 (dd, J=9.5, 3.9 Hz, 1H), 4.46-4.41 (m, 1H), 4.38 (d, J=17.3 Hz, 1H), 4.17 (t, J=10.6 Hz, 1H), 4.08-4.00 (m, 2H), 3.92-3.88 (m, 1H), 3.09 (dt, J=7.0, 4.5 Hz, 2H), 2.91-2.87 (m, 1H), 2.79-2.73 (m, 1H), 2.57-2.53 (m, 2H), 2.51-2.46 (m, 2H), 2.27-2.23 (m, 1H), 2.21-2.11 (m, 1H), 1.80-1.70 (m, 4H), 1.62-1.57 (m, 2H). HRMS (ESI) m/z: calcd for C37H37ClFN6O6S+ [M+H]+, 747.2162; found, 747.2161.
According to Scheme 9, to a solution of Dacomitinib derivative B (20 mg, 0.036 mmol) dissolved in 2 mL of DMF was sequentially added SIAIS255121 (17.5 mg, 0.0432 mmol), NaI (10.8 mg, 0.072 mmol), and K2CO3 (10 mg, 0.072 mmol), and the reaction mixture was heated to 50° C. and reacted overnight. The reaction was quenched with 0.10 mL of water. The resulting mixture was subjected to preparative HPLC (eluent (v/v): acetonitrile/(water+0.05% HCl)=10%-100%) for separation, and the collected fractions were rotary evaporated under reduced pressure to remove the acetonitrile and most of the water. The resulting residue was lyophilized to give the final target compound (SIAIS262050). (yellow solid, 12.1 mg, yield 39%)1H NMR (500 MHz, MeOD) δ 9.26 (s, 1H), 8.74 (s, 1H), 7.91 (dd, J=6.6, 2.6 Hz, 1H), 7.74 (d, J=7.4 Hz, 1H), 7.67-7.62 (m, 2H), 7.50 (t, J=7.6 Hz, 1H), 7.36 (t, J=8.9 Hz, 1H), 7.32 (s, 1H), 7.11-7.02 (m, 1H), 6.86 (d, J=15.5 Hz, 1H), 5.20 (dd, J=13.2, 5.2 Hz, 1H), 4.57 (d, J=17.6 Hz, 1H), 4.51 (d, J=17.6 Hz, 1H), 4.19 (s, 3H), 4.06 (s, 2H), 3.72 (s, 5H), 3.60-3.37 (m, 8H), 3.16 (d, J=13.9 Hz, 2H), 2.97-2.88 (m, 1H), 2.77 (d, J=15.6 Hz, 1H), 2.63-2.58 (m, 3H), 2.38 (s, 2H), 2.24-2.16 (m, 1H), 2.11-1.99 (m, 2H), 1.77-1.71 (m, 2H). HRMS (ESI) m/z: calcd for C46H50ClFN9O5+ [M+H]+, 862.3602; found, 862.3601.
Referring to the method of example 102, the target compound (SIAIS262051) was prepared by using Dacomitinib derivative B and intermediate LM (SIAIS255119). (yellow solid, 12.8 mg, yield 41%) 1H NMR (500 MHz, MeOD) δ 9.27 (s, 1H), 8.76 (s, 1H), 7.93 (dd, J=6.6, 2.6 Hz, 1H), 7.75 (d, J=7.4 Hz, 1H), 7.68-7.62 (m, 2H), 7.51 (t, J=7.6 Hz, 1H), 7.38 (t, J=8.9 Hz, 1H), 7.33 (s, 1H), 7.10-7.02 (m, 1H), 6.87 (d, J=15.5 Hz, 1H), 5.21 (dd, J=13.2, 5.2 Hz, 1H), 4.58 (d, J=17.6 Hz, 1H), 4.52 (d, J=17.6 Hz, 1H), 4.19 (s, 3H), 4.08 (s, 2H), 3.74 (s, 5H), 3.61-3.37 (m, 8H), 3.18 (d, J=13.9 Hz, 2H), 2.98-2.88 (m, 1H), 2.79 (d, J=15.6 Hz, 1H), 2.63-2.59 (m, 3H), 2.39 (s, 2H), 2.25-2.16 (m, 1H), 2.12-1.99 (m, 4H), 1.79-1.71 (m, 2H). HRMS (ESI) m/z: calcd for C47H52ClFN9O5+ [M+H]+, 876.3758; found, 876.3755.
Referring to the method of example 102, the target compound (SIAIS262089) was prepared by using Gefitinib derivative B and intermediate LM (SIAIS255121). (yellow solid, 13.1 mg, yield 41%) 1H NMR (500 MHz, MeOD) δ 8.74 (s, 1H), 8.12 (s, 1H), 7.98 (s, 1H), 7.77 (d, J=8.0 Hz, 1H), 7.70 (s, 1H), 7.65 (d, J=7.2 Hz, 1H), 7.53 (d, J=7.5 Hz, 1H), 7.37 (t, J=8.5 Hz, 1H), 7.26 (s, 1H), 5.20 (d, J=7.1 Hz, 1H), 4.56 (s, 1H), 4.51 (d, J=17.5 Hz, 1H), 4.42 (s, 2H), 4.10 (s, 3H), 3.83 (d, J=11.2 Hz, 2H), 3.75-3.32 (m, 13H), 3.15 (d, J=12.9 Hz, 2H), 2.93 (s, 1H), 2.81-2.77 (m, 1H), 2.68 (s, 2H), 2.55 (s, 1H), 2.44 (s, 2H), 2.28 (s, 2H), 2.18-2.13 (m, 3H), 2.02 (s, 2H). HRMS (ESI) m/z: calcd for C45H51ClFN8O5+ [M+H]+, 837.3649; found, 837.3644.
Referring to the method of example 102, the target compound (SIAIS262090) was prepared by using Gefitinib derivative B and intermediate LM (SIAIS255119). (yellow solid, 13.1 mg, yield 41%) 1H NMR (500 MHz, MeOD) δ 8.74 (s, 1H), 8.11 (s, 1H), 7.97 (dd, J=6.6, 2.6 Hz, 1H), 7.76 (d, J=7.9 Hz, 1H), 7.71 (dd, J=7.9, 3.7 Hz, 1H), 7.64 (d, J=6.9 Hz, 1H), 7.51 (t, J=7.7 Hz, 1H), 7.37 (t, J=8.9 Hz, 1H), 7.26 (s, 1H), 5.20 (dd, J=13.3, 5.2 Hz, 1H), 4.56 (d, J=17.4 Hz, 1H), 4.50 (d, J=17.4 Hz, 1H), 4.42 (t, J=5.7 Hz, 2H), 4.10 (s, 3H), 3.85 (d, J=11.7 Hz, 2H), 3.55-3.40 (m, 5H), 3.28-3.04 (m, 8H), 2.95-2.90 (m, 1H), 2.79 (d, J=15.6 Hz, 1H), 2.64-2.51 (m, 3H), 2.43 (s, 2H), 2.29 (d, J=7.1 Hz, 2H), 2.20 (s, 1H), 1.99 (s, 4H), 1.75 (d, J=7.7 Hz, 2H), 1.60 (s, 2H). HRMS (ESI) m/z: calcd for C46H53ClFN8O5+ [M+H]+, 851.3806; found, 851.3802.
Referring to the method of example 102, the target compound (SIAIS262065) was prepared by using Dacomitinib derivative B and intermediate LM (SIAIS1216133). (yellow solid, 11.2 mg, yield 45%). 1H NMR (500 MHz, MeOD) δ 9.27 (s, 1H), 8.76 (s, 1H), 7.93 (dd, J=6.6, 2.6 Hz, 1H), 7.66 (dt, J=8.7, 4.3 Hz, 3H), 7.54 (t, J=7.7 Hz, 1H), 7.38 (t, J=8.9 Hz, 1H), 7.33 (s, 1H), 7.06 (dt, J=14.7, 7.2 Hz, 1H), 6.88 (d, J=15.4 Hz, 1H), 5.18 (dd, J=13.4, 5.2 Hz, 1H), 4.47 (d, J=17.3 Hz, 1H), 4.45-4.39 (m, 1H), 4.18 (d, J=5.9 Hz, 3H), 4.08 (d, J=6.0 Hz, 2H), 3.76 (s, 2H), 3.75-3.35 (m, 8H), 3.18 (s, 4H), 3.08 (dq, J=13.2, 6.3 Hz, 3H), 2.97-2.88 (m, 1H), 2.80 (ddd, J=17.7, 4.6, 2.4 Hz, 1H), 2.60-2.51 (m, 1H), 2.43 (s, 2H), 2.23-2.07 (m, 3H), 1.79-1.66 (m, 4H), 1.57-1.51 (m, 2H), 1.42 (dd, J=14.7, 7.3 Hz, 2H). HRMS (ESI) m/z: calcd for C47H56ClFN9O5S+ [M+H]+, 912.3792; found, 912.3790.
Referring to the method of example 102, the target compound (SIAIS262072) was prepared by using Dacomitinib derivative B and intermediate LM (SIAIS264018). (yellow solid, 10.8 mg, yield 44%). 1H NMR (500 MHz, MeOD) δ 9.27 (s, 1H), 8.75 (s, 1H), 7.92 (dd, J=6.6, 2.6 Hz, 1H), 7.67-7.63 (m, 1H), 7.58-7.54 (m, 1H), 7.38 (t, J=8.9 Hz, 1H), 7.33 (s, 1H), 7.05 (q, J=8.3 Hz, 3H), 6.87 (d, J=15.1 Hz, 1H), 5.07 (dd, J=12.6, 5.7 Hz, 1H), 4.18 (s, 3H), 4.07 (s, 2H), 3.74 (s, 4H), 3.39 (dd, J=26.0, 19.4 Hz, 5H), 3.19 (s, 4H), 2.85 (dd, J=13.0, 4.4 Hz, 1H), 2.78-2.69 (m, 2H), 2.38 (s, 2H), 2.13-2.03 (m, 3H), 1.80 (s, 2H), 1.71 (dd, J=14.0, 6.7 Hz, 2H), 1.49 (d, J=9.3 Hz, 4H). HRMS (ESI) m/z: calcd for C47H55ClFN10O6+ [M+H]+, 909.3973; found, 909.3971.
Referring to the method of example 1, the target compound (SIAIS262121) was prepared by using Caneritinib derivative A and intermediate LM (SIAIS171090). (yellow solid, 6.8 mg, yield 40%). 1H NMR (500 MHz, MeOD) δ 9.14 (s, 1H), 8.75 (s, 1H), 7.95 (dd, J=6.6, 2.5 Hz, 1H), 7.83 (d, J=7.7 Hz, 1H), 7.76 (d, J=7.6 Hz, 1H), 7.68-7.63 (m, 1H), 7.58 (t, J=7.6 Hz, 1H), 7.39-7.33 (m, 2H), 6.79 (dd, J=17.0, 10.3 Hz, 1H), 6.51 (d, J=16.9 Hz, 1H), 5.90 (d, J=11.0 Hz, 1H), 5.18 (dd, J=13.3, 5.1 Hz, 1H), 4.55 (s, 1H), 4.48 (dd, J=14.6, 8.9 Hz, 4H), 4.02 (d, J=17.2 Hz, 2H), 3.60 (s, 4H), 3.45 (s, 4H), 2.93-2.88 (m, 1H), 2.78 (s, 1H), 2.58-2.51 (m, 1H), 2.46 (s, 2H), 2.19 (d, J=7.7 Hz, 1H). HRMS (ESI) m/z: calcd for C39H39ClFN8O6S+ [M+H]+, 801.2380; found, 801.2381.
Referring to the method of example 1, the target compound (SIAIS262122) was prepared by using Caneritinib derivative A and intermediate LM (SIAIS171079). (yellow solid, 7.1 mg, yield 40%). 1H NMR (500 MHz, MeOD) δ 9.11 (s, 1H), 8.74 (s, 1H), 7.95 (dd, J=6.6, 2.6 Hz, 1H), 7.69-7.64 (m, 3H), 7.53 (d, J=7.6 Hz, 1H), 7.37 (t, J=8.9 Hz, 1H), 7.34 (s, 1H), 6.75 (dd, J=16.9, 10.3 Hz, 1H), 6.51 (dd, J=16.9, 1.5 Hz, 1H), 5.90 (dd, J=10.2, 1.5 Hz, 1H), 5.15 (dd, J=13.4, 5.1 Hz, 1H), 4.48 (dd, J=11.5, 5.8 Hz, 3H), 4.41 (d, J=17.3 Hz, 1H), 3.64 (s, 4H), 3.45 (s, 2H), 3.23-2.97 (m, 6H), 2.93-2.86 (m, 1H), 2.79 (dd, J=10.0, 7.6 Hz, 1H), 2.54 (dd, J=13.1, 4.8 Hz, 1H), 2.46 (t, J=7.1 Hz, 4H), 2.21-2.16 (m, 1H), 1.74 (dd, J=23.6, 6.9 Hz, 4H). HRMS (ESI) m/z: calcd for C42H45ClFN8O6S+ [M+H]+, 843.2850; found, 843.2853.
Referring to the method of example 1, the target compound (SIAIS262123) was prepared by using Caneritinib derivative A and intermediate LM (SIAIS171091). (yellow solid, 8.2 mg, yield 46%). 1H NMR (500 MHz, MeOD) δ 9.14 (s, 1H), 8.75 (s, 1H), 7.95 (dd, J=6.6, 2.6 Hz, 1H), 7.69-7.63 (m, 3H), 7.54 (t, J=7.6 Hz, 1H), 7.38 (t, J=8.9 Hz, 1H), 7.35 (s, 1H), 6.78 (dd, J=17.0, 10.2 Hz, 1H), 6.51 (d, J=16.8 Hz, 1H), 5.91 (d, J=10.4 Hz, 1H), 5.18-5.13 (m, 1H), 4.47 (dd, J=11.2, 5.3 Hz, 3H), 4.42 (d, J=17.3 Hz, 1H), 3.63 (d, J=30.6 Hz, 4H), 3.46 (s, 2H), 3.09 (ddd, J=22.7, 18.4, 16.1 Hz, 6H), 2.95-2.85 (m, 1H), 2.78 (d, J=15.2 Hz, 1H), 2.55 (dd, J=13.2, 4.7 Hz, 1H), 2.50 (d, J=9.4 Hz, 2H), 2.40 (t, J=7.2 Hz, 2H), 2.19 (d, J=7.9 Hz, 1H), 1.68 (dd, J=14.5, 7.1 Hz, 2H), 1.61 (dd, J=14.6, 7.4 Hz, 2H), 1.52 (d, J=7.3 Hz, 2H). HRMS (ESI) m/z: calcd for C43H47ClFN8O6S+ [M+H]+, 857.3006; found, 857.3002.
Referring to the method of example 1, the target compound (SIAIS262124) was prepared by using Caneritinib derivative A and intermediate LM (SIAIS151025). (yellow solid, 7.3 mg, yield 44%). 1H NMR (500 MHz, MeOD) δ 9.10 (s, 1H), 8.73 (s, 1H), 7.95 (dd, J=6.5, 2.5 Hz, 1H), 7.68-7.63 (m, 1H), 7.57 (d, J=8.5 Hz, 1H), 7.39-7.33 (m, 2H), 7.11 (d, J=7.0 Hz, 1H), 7.03 (d, J=8.4 Hz, 1H), 6.76 (dd, J=17.0, 10.3 Hz, 1H), 6.51 (dd, J=16.9, 1.5 Hz, 1H), 5.91 (dd, J=10.3, 1.5 Hz, 1H), 5.10-5.06 (m, 1H), 4.48 (t, J=5.8 Hz, 2H), 4.29 (s, 2H), 3.62 (d, J=21.2 Hz, 4H), 3.47 (d, J=18.1 Hz, 4H), 3.17 (s, 2H), 2.84 (dd, J=13.0, 7.8 Hz, 1H), 2.74 (dd, J=18.5, 5.6 Hz, 3H), 2.50-2.47 (m, 1H), 2.13 (d, J=5.1 Hz, 1H). HRMS (ESI) m/z: calcd for C39H38ClFN9O7+ [M+H]+, 798.2561; found, 798.2564.
Referring to the method of example 1, the target compound (SIAIS262125) was prepared by using Caneritinib derivative A and intermediate LM (SIAIS151027). (yellow solid, 7.9 mg, yield 44%). 1H NMR (500 MHz, MeOD) δ 9.08 (s, 1H), 8.73 (s, 1H), 7.95 (d, J=3.9 Hz, 1H), 7.65 (s, 1H), 7.55 (d, J=8.2 Hz, 1H), 7.37 (t, J=8.9 Hz, 1H), 7.33 (s, 1H), 7.08-7.02 (m, 2H), 6.72 (dd, J=16.9, 10.7 Hz, 1H), 6.51 (d, J=16.9 Hz, 1H), 5.90 (d, J=10.6 Hz, 1H), 5.05 (d, J=11.9 Hz, 1H), 4.46 (s, 2H), 4.28-4.08 (m, 2H), 3.58 (s, 4H), 3.45 (s, 4H), 3.14-3.02 (m, 2H), 2.84 (d, J=13.8 Hz, 1H), 2.72-2.68 (m, 2H), 2.48 (d, J=6.9 Hz, 4H), 2.11 (s, 1H), 1.70 (s, 4H), 1.50 (s, 2H). HRMS (ESI) m/z: calcd for C43H46ClFN9O7+ [M+H]+, 854.3187; found, 854.3182.
Referring to the method of example 1, the target compound (SIAIS262126) was prepared by using Caneritinib derivative A and intermediate LM (SIAIS151086). (yellow solid, 7.6 mg, yield 42%). 1H NMR (500 MHz, MeOD) δ 9.12 (s, 1H), 8.74 (s, 1H), 7.95 (dd, J=6.6, 2.4 Hz, 1H), 7.68-7.63 (m, 1H), 7.59-7.53 (m, 1H), 7.37 (t, J=8.8 Hz, 1H), 7.34 (s, 1H), 7.05 (t, J=8.0 Hz, 2H), 6.75 (dd, J=16.9, 10.2 Hz, 1H), 6.51 (d, J=17.7 Hz, 1H), 5.91 (d, J=10.5 Hz, 1H), 5.07-5.03 (m, 1H), 4.47 (t, J=5.5 Hz, 2H), 3.60 (s, 4H), 3.45 (s, 4H), 3.17 (s, 4H), 2.85 (d, J=13.4 Hz, 1H), 2.72 (t, J=13.1 Hz, 2H), 2.52-2.42 (m, 4H), 2.13-2.08 (m, 1H), 1.73-1.62 (m, 4H), 1.46 (s, 4H). HRMS (ESI) m/z: calcd for C44H48ClFN9O7+ [M+H]+, 868.3344; found, 868.3341.
Referring to the method of example 1, the target compound (SIAIS262127) was prepared by using Caneritinib derivative A and intermediate LM (SIAIS151004). (yellow solid, 8.5 mg, yield 45%). 1H NMR (500 MHz, MeOD) δ 9.06 (s, 1H), 8.70 (s, 1H), 7.95 (dd, J=6.7, 2.6 Hz, 1H), 7.67 (dd, J=7.2, 4.5 Hz, 1H), 7.48-7.43 (m, 1H), 7.38 (t, J=8.9 Hz, 1H), 7.27 (s, 1H), 7.00 (d, J=8.5 Hz, 1H), 6.91 (d, J=7.2 Hz, 1H), 6.73 (dd, J=16.9, 10.3 Hz, 1H), 6.55-6.48 (m, 1H), 5.93-5.89 (m, 1H), 5.06 (dd, J=12.8, 5.5 Hz, 3H), 4.38 (s, 2H), 3.81-3.56 (m, 12H), 3.44 (d, J=11.5 Hz, 8H), 2.89-2.81 (m, 2H), 2.75 (d, J=7.4 Hz, 2H), 2.67 (d, J=13.4 Hz, 1H), 2.44 (s, 2H), 2.11 (s, 1H). HRMS (ESI) m/z: calcd for C44H48ClFN9O9+ [M+H]+, 900.3242; found, 900.3241.
Referring to the method of example 1, the target compound (SIAIS262128) was prepared by using Caneritinib derivative A and intermediate LM (SIAIS074012). (yellow solid, 10.3 mg, yield 48%). 1H NMR (500 MHz, MeOD) δ 9.40 (d, J=10.0 Hz, 1H), 9.17 (s, 1H), 8.76 (s, 1H), 7.94 (dd, J=6.6, 2.6 Hz, 1H), 7.68-7.63 (m, 1H), 7.51 (t, J=7.7 Hz, 2H), 7.46 (d, J=8.3 Hz, 2H), 7.39-7.35 (m, 2H), 6.85 (dd, J=16.9, 10.3 Hz, 1H), 6.51 (dd, J=16.9, 1.5 Hz, 1H), 5.90 (dd, J=10.3, 1.5 Hz, 1H), 4.58 (dd, J=13.6, 5.6 Hz, 2H), 4.50 (dd, J=12.6, 7.2 Hz, 4H), 4.41-4.36 (m, 1H), 4.22 (s, 1H), 3.91 (d, J=11.0 Hz, 1H), 3.81 (dd, J=11.0, 3.8 Hz, 1H), 3.66 (d, J=35.5 Hz, 3H), 3.51 (s, 2H), 3.18 (d, J=12.4 Hz, 2H), 2.55-2.49 (m, 5H), 2.46 (t, J=7.5 Hz, 2H), 2.29-2.23 (m, 3H), 2.11-2.05 (m, 1H), 1.66-1.59 (m, 4H), 1.41-1.35 (m, 4H), 1.04 (s, 9H). HRMS (ESI) m/z: calcd for C51H61ClFN10O7S+ [M+H]+, 1011.4112; found, 1011.4114.
Referring to the method of example 102, the target compound (SIAIS262131) was prepared by using Caneritinib derivative A and intermediate LM (SIAIS255119). (yellow solid, 9.4 mg, yield 38%). 1H NMR (500 MHz, MeOD) δ 9.18 (s, 1H), 8.76 (s, 1H), 7.94 (dd, J=6.6, 2.6 Hz, 1H), 7.76 (d, J=7.1 Hz, 1H), 7.68-7.62 (m, 2H), 7.51 (t, J=7.6 Hz, 1H), 7.38 (t, J=8.9 Hz, 1H), 7.35 (s, 1H), 6.81 (dd, J=16.8, 10.4 Hz, 1H), 6.51 (dd, J=16.9, 1.5 Hz, 1H), 5.90 (dd, J=10.2, 1.5 Hz, 1H), 5.21 (dd, J=13.3, 5.2 Hz, 1H), 4.56 (s, 1H), 4.52 (d, J=12.1 Hz, 1H), 4.49 (d, J=5.9 Hz, 2H), 3.75 (s, 4H), 3.65-3.38 (m, 8H), 2.95-2.87 (m, 1H), 2.80 (dd, J=15.3, 12.9 Hz, 1H), 2.63 (t, J=6.7 Hz, 2H), 2.60-2.53 (m, 1H), 2.46 (s, 2H), 2.22-2.15 (m, 1H), 2.03 (d, J=7.9 Hz, 2H), 1.76 (dd, J=14.7, 7.3 Hz, 2H). HRMS (ESI) m/z: calcd for C43H45ClFN8O5+ [M+H]+, 807.3180; found, 807.3177.
Referring to the method of example 102, the target compound (SIAIS262182) was prepared by using Caneritinib derivative A and intermediate LM (SIAIS264018). (yellow solid, 12.1 mg, yield 46%). 1H NMR (500 MHz, DMSO) δ 11.09 (s, 1H), 9.94 (s, 1H), 9.16 (s, 1H), 8.87 (s, 1H), 7.99 (d, J=5.9 Hz, 1H), 7.68 (dd, J=7.5, 4.2 Hz, 1H), 7.59 (dd, J=8.4, 7.2 Hz, 1H), 7.54 (t, J=9.0 Hz, 1H), 7.43 (s, 1H), 7.11 (d, J=8.6 Hz, 1H), 7.03 (d, J=7.0 Hz, 1H), 6.94 (s, 1H), 6.55 (s, 1H), 6.37 (d, J=16.9 Hz, 1H), 5.86 (d, J=11.1 Hz, 1H), 5.05 (dd, J=12.8, 5.4 Hz, 1H), 4.38 (s, 2H), 3.73 (d, J=47.0 Hz, 6H), 3.22 (d, J=102.4 Hz, 10H), 2.89-2.83 (m, 1H), 2.64-2.52 (m, 2H), 2.36 (s, 2H), 2.07-1.98 (m, 1H), 1.72 (s, 2H), 1.67-1.56 (m, 2H), 1.38 (s, 4H). HRMS (ESI) m/z: calcd for C43H48ClFN9O6+ [M+H]+, 840.3395; found, 840.3392.
Referring to the method of example 1, the target compound (SIAIS262174) was prepared by using Caneritinib derivative B and intermediate LM (SIAIS171090). (yellow solid, 6.8 mg, yield 44%). 1H NMR (500 MHz, MeOD) δ 9.17 (s, 1H), 8.75 (s, 1H), 7.93 (dd, J=6.6, 2.6 Hz, 1H), 7.69-7.64 (m, 2H), 7.53 (t, J=7.6 Hz, 1H), 7.34 (dd, J=11.7, 6.0 Hz, 2H), 6.85-6.82 (m, 1H), 6.50 (d, J=16.9 Hz, 1H), 5.91 (d, J=10.3 Hz, 1H), 5.16 (dd, J=13.4, 5.1 Hz, 1H), 4.46 (dd, J=11.5, 5.9 Hz, 3H), 4.41 (d, J=17.3 Hz, 1H), 3.87 (d, J=12.0 Hz, 2H), 3.66 (s, 2H), 3.56-3.43 (m, 4H), 3.25 (t, J=12.5 Hz, 4H), 3.17-3.06 (m, 3H), 2.95-2.85 (m, 1H), 2.76 (d, J=15.5 Hz, 1H), 2.60-2.44 (m, 7H), 2.26 (d, J=12.5 Hz, 2H), 2.21-2.16 (m, 1H). HRMS (ESI) m/z: calcd for C44H48ClFN9O6S+ [M+H]+, 884.3115; found, 884.3119.
Referring to the method of example 1, the target compound (SIAIS262175) was prepared by using Caneritinib derivative B and intermediate LM (SIAIS171079). (yellow solid, 7.1 mg, yield 44%). 1H NMR (500 MHz, MeOD) δ 9.18 (s, 1H), 8.76 (s, 1H), 7.94 (dd, J=6.6, 2.6 Hz, 1H), 7.69-7.65 (m, 2H), 7.54 (t, J=7.6 Hz, 1H), 7.37 (dd, J=11.7, 6.0 Hz, 2H), 6.85 (dd, J=16.8, 10.3 Hz, 1H), 6.51 (d, J=16.9 Hz, 1H), 5.91 (d, J=10.3 Hz, 1H), 5.17 (dd, J=13.4, 5.1 Hz, 1H), 4.48 (dd, J=11.5, 5.9 Hz, 3H), 4.42 (d, J=17.3 Hz, 1H), 3.87 (d, J=12.0 Hz, 2H), 3.68 (s, 2H), 3.56-3.40 (m, 4H), 3.24 (t, J=12.5 Hz, 4H), 3.17-3.04 (m, 3H), 2.97-2.87 (m, 1H), 2.79 (d, J=15.5 Hz, 1H), 2.60-2.44 (m, 7H), 2.28 (d, J=12.5 Hz, 2H), 2.22-2.16 (m, 1H), 1.83-1.68 (m, 4H), 1.45-1.32 (m, 2H). HRMS (ESI) m/z: calcd for C47H54ClFN9O6S+ [M+H]+, 926.3585; found, 926.3583.
Referring to the method of example 1, the target compound (SIAIS262176) was prepared by using Caneritinib derivative B and intermediate LM (SIAIS171091). (yellow solid, 7.5 mg, yield 45%). 1H NMR (500 MHz, MeOD) δ 9.17 (s, 1H), 8.76 (s, 1H), 7.94 (dd, J=6.6, 2.6 Hz, 1H), 7.67 (dd, J=7.7, 2.9 Hz, 2H), 7.54 (t, J=7.6 Hz, 1H), 7.38 (t, J=8.9 Hz, 1H), 7.34 (s, 1H), 6.82 (dd, J=16.9, 10.3 Hz, 1H), 6.52 (dd, J=16.9, 1.4 Hz, 1H), 5.95-5.88 (m, 1H), 5.18 (dd, J=13.1, 5.0 Hz, 1H), 4.48 (dd, J=11.5, 6.0 Hz, 3H), 4.42 (d, J=17.3 Hz, 1H), 3.86 (d, J=11.1 Hz, 2H), 3.44 (dd, J=33.3, 26.1 Hz, 8H), 3.30-3.14 (m, 4H), 3.08 (ddd, J=19.6, 13.2, 6.3 Hz, 3H), 2.91 (dd, J=21.7, 9.2 Hz, 1H), 2.79 (d, J=15.7 Hz, 1H), 2.60-2.44 (m, 5H), 2.39 (t, J=7.0 Hz, 2H), 2.19 (dd, J=15.9, 10.9 Hz, 3H), 1.67 (dd, J=15.0, 7.2 Hz, 2H), 1.61 (dd, J=14.8, 7.3 Hz, 2H), 1.56-1.49 (m, 2H). HRMS (ESI) m/z: calcd for C48H56ClFN9O6S+ [M+H]+, 940.3741; found, 940.3739.
Referring to the method of example 1, the target compound (SIAIS262177) was prepared by using Caneritinib derivative B and intermediate LM (SIAIS151025). (yellow solid, 6.4 mg, yield 41%). 1H NMR (500 MHz, MeOD) δ 9.17 (s, 1H), 8.75 (s, 1H), 7.94 (dd, J=6.6, 2.6 Hz, 1H), 7.69-7.63 (m, 1H), 7.62-7.54 (m, 1H), 7.38 (t, J=8.9 Hz, 1H), 7.34 (s, 1H), 7.11 (d, J=7.1 Hz, 1H), 7.03 (d, J=8.5 Hz, 1H), 6.81 (dd, J=16.9, 10.3 Hz, 1H), 6.52 (d, J=16.9 Hz, 1H), 5.91 (d, J=11.6 Hz, 1H), 5.08 (dd, J=12.6, 5.5 Hz, 1H), 4.49 (t, J=5.5 Hz, 2H), 4.29 (s, 2H), 3.88 (d, J=10.5 Hz, 2H), 3.62-3.58 (m, 8H), 3.29-2.95 (m, 5H), 2.86 (dd, J=13.5, 5.7 Hz, 1H), 2.80-2.68 (m, 2H), 2.51 (s, 4H), 2.25 (s, 2H), 2.16-2.09 (m, 1H). HRMS (ESI) m/z: calcd for C44H47ClFN10O7+ [M+H]+, 881.3296; found, 881.3296.
Referring to the method of example 1, the target compound (SIAIS262178) was prepared by using Caneritinib derivative B and intermediate LM (SIAIS151027). (yellow solid, 7.8 mg, yield 47%). 1H NMR (500 MHz, MeOD) δ 9.17 (s, 1H), 8.76 (s, 1H), 7.94 (dd, J=6.6, 2.6 Hz, 1H), 7.69-7.62 (m, 1H), 7.59-7.54 (m, 1H), 7.38 (t, J=8.9 Hz, 1H), 7.35 (s, 1H), 7.06 (t, J=7.7 Hz, 1H), 6.83 (dd, J=16.8, 10.4 Hz, 1H), 6.52 (d, J=16.9 Hz, 1H), 5.91 (d, J=11.8 Hz, 1H), 5.07 (dd, J=12.6, 5.5 Hz, 1H), 4.49 (s, 2H), 3.87 (s, 2H), 3.71-3.35 (m, 12H), 3.21 (d, J=10.6 Hz, 5H), 2.85 (d, J=8.6 Hz, 1H), 2.73 (t, J=14.4 Hz, 2H), 2.55-2.45 (m, 4H), 2.27 (s, 2H), 2.12 (s, 1H), 1.74-1.67 (m, 2H), 1.51 (d, J=7.3 Hz, 2H), 1.41-1.33 (m, 2H). HRMS (ESI) m/z: calcd for C48H55ClFN10O7+ [M+H]+, 937.3922; found, 937.3926.
Referring to the method of example 1, the target compound (SIAIS262179) was prepared by using Caneritinib derivative B and intermediate LM (SIAIS151086). (yellow solid, 8.2 mg, yield 49%). 1H NMR (500 MHz, MeOD) δ 9.17 (s, 1H), 8.75 (s, 1H), 7.97-7.92 (m, 1H), 7.68-7.61 (m, 1H), 7.59-7.53 (m, 1H), 7.37 (dd, J=15.8, 6.9 Hz, 2H), 7.07-7.04 (m, 1H), 6.82 (s, 1H), 6.51 (d, J=17.0 Hz, 1H), 5.91 (dd, J=10.3, 1.5 Hz, 1H), 5.07 (dd, J=12.1, 6.0 Hz, 1H), 4.49 (s, 2H), 3.84 (s, 2H), 3.72-3.32 (m, 12H), 3.18 (d, J=15.7 Hz, 5H), 2.85 (dd, J=13.3, 4.8 Hz, 1H), 2.78-2.67 (m, 2H), 2.47 (d, J=23.2 Hz, 4H), 2.31-2.17 (m, 2H), 2.15-2.08 (m, 1H), 1.73-1.60 (m, 4H), 1.45 (d, J=13.6 Hz, 4H). HRMS (ESI) m/z: calcd for C49H57ClFN10O7+[M+H]+, 951.4079; found, 951.4075.
Referring to the method of example 102, the target compound (SIAIS262180) was prepared by using Caneritinib derivative B and intermediate LM (SIAIS1216133). (yellow solid, 11.2 mg, yield 46%). 1H NMR (500 MHz, MeOD) δ 9.17 (s, 1H), 8.76 (s, 1H), 7.94 (dd, J=6.6, 2.6 Hz, 1H), 7.67-7.65 (m, 2H), 7.54 (t, J=7.6 Hz, 1H), 7.38 (dd, J=12.3, 5.4 Hz, 2H), 6.84 (dd, J=16.9, 10.3 Hz, 1H), 6.52 (dd, J=16.9, 1.3 Hz, 1H), 5.91 (dd, J=10.3, 1.4 Hz, 1H), 5.18 (dd, J=13.3, 5.2 Hz, 1H), 4.48 (dd, J=13.2, 7.4 Hz, 3H), 4.44-4.39 (m, 1H), 3.83 (d, J=12.5 Hz, 2H), 3.76-3.34 (m, 10H), 3.19 (t, J=12.2 Hz, 4H), 3.09-3.04 (m, 3H), 2.96-2.88 (m, 1H), 2.84-2.76 (m, 1H), 2.55-2.51 (m, 3H), 2.40 (d, J=12.8 Hz, 2H), 2.19-2.15 (m, 3H), 1.78-1.67 (m, 4H), 1.53 (d, J=7.3 Hz, 2H), 1.41 (dt, J=13.8, 7.2 Hz, 2H). HRMS (ESI) m/z: calcd for C48H55ClFN9O5S+ [M+H]+, 926.3949; found, 926.3953.
Referring to the method of example 102, the target compound (SIAIS262183) was prepared by using Caneritinib derivative B and intermediate LM (SIAIS264018). (yellow solid, 12.2 mg, yield 50%). 1H NMR (500 MHz, MeOD) δ 9.17 (s, 1H), 8.76 (s, 1H), 7.94 (dd, J=6.5, 2.5 Hz, 1H), 7.66-7.61 (m, 1H), 7.56 (dd, J=8.5, 7.1 Hz, 1H), 7.40-7.34 (m, 2H), 7.06 (t, J=7.4 Hz, 2H), 6.84 (dd, J=16.9, 10.0 Hz, 1H), 6.52 (d, J=16.9 Hz, 1H), 5.91 (d, J=11.7 Hz, 1H), 5.07 (dd, J=12.6, 5.5 Hz, 1H), 4.48 (t, J=5.6 Hz, 2H), 3.80 (s, 2H), 3.43-3.41 (m, 8H), 3.18 (d, J=13.1 Hz, 4H), 2.88-2.83 (m, 1H), 2.77-2.69 (m, 2H), 2.50 (s, 2H), 2.34 (s, 2H), 2.16-2.08 (m, 3H), 1.80 (s, 2H), 1.76-1.70 (m, 2H), 1.50 (d, J=10.7 Hz, 4H). HRMS (ESI) m/z: calcd for C48H57ClFN10O6+ [M+H]+, 923.4130; found, 923.4143.
Referring to the method of example 1, the target compound (SIAIS293047) was prepared by using Gefitinib derivative C and intermediate LM (SIAIS171123). (yellow solid, 7.6 mg, yield 45%). 1H NMR (500 MHz, MeOD) δ 8.73 (s, 1H), 8.31 (s, 1H), 8.00 (dd, J=6.6, 2.6 Hz, 1H), 7.78 (dd, J=7.8, 0.8 Hz, 1H), 7.72 (s, 1H), 7.66 (s, 1H), 7.56 (t, J=7.7 Hz, 1H), 7.36 (t, J=8.9 Hz, 1H), 7.28 (s, 1H), 5.17 (d, J=8.4 Hz, 1H), 5.01 (s, 1H), 4.53 (d, J=17.4 Hz, 1H), 4.47 (d, J=17.5 Hz, 1H), 4.11 (d, J=9.7 Hz, 3H), 3.90 (q, J=15.5 Hz, 2H), 3.58-3.50 (m, 2H), 3.49-3.33 (m, 4H), 3.29-3.21 (m, 2H), 2.90 (ddd, J=17.0, 12.7, 4.5 Hz, 1H), 2.83-2.74 (m, 1H), 2.52 (dt, J=13.1, 8.4 Hz, 1H), 2.39-2.17 (m, 4H), 2.04 (s, 1H). HRMS (ESI) m/z: calcd for C37H38ClFN7O6S+ [M+H]+, 762.2271; found, 762.2266.
Referring to the method of example 1, the target compound (SIAIS293048) was prepared by using Gefitinib derivative C and intermediate LM (SIAIS171124). (yellow solid, 8.1 mg, yield 47%). 1H NMR (500 MHz, MeOD) δ 8.74 (s, 1H), 8.33 (s, 1H), 7.99 (dd, J=6.6, 2.6 Hz, 1H), 7.72 (ddd, J=8.9, 4.1, 2.7 Hz, 1H), 7.68 (d, J=7.6 Hz, 1H), 7.64 (d, J=7.3 Hz, 1H), 7.52 (t, J=7.7 Hz, 1H), 7.36 (t, J=8.9 Hz, 1H), 7.28 (s, 1H), 5.17 (d, J=9.5 Hz, 1H), 5.04 (s, 1H), 4.50 (s, 1H), 4.45 (d, J=17.4 Hz, 1H), 4.09 (s, 3H), 3.99 (s, 2H), 3.67-3.34 (m, 6H), 3.20 (ddd, J=15.2, 14.2, 9.6 Hz, 2H), 3.10 (t, J=7.2 Hz, 2H), 2.95-2.85 (m, 1H), 2.82-2.75 (m, 1H), 2.53 (ddd, J=26.5, 13.3, 4.7 Hz, 1H), 2.32 (s, 2H), 2.22-2.16 (m, 1H), 1.93-1.83 (m, 2H). HRMS (ESI) m/z: calcd for C38H40ClFN7O6S+ [M+H]+, 776.2428; found, 776.2424.
Referring to the method of example 1, the target compound (SIAIS293049) was prepared by using Gefitinib derivative C and intermediate LM (SIAIS171131). (yellow solid, 8.4 mg, yield 48%). 1H NMR (500 MHz, MeOD) δ 8.74 (s, 1H), 8.31 (s, 1H), 7.99 (dd, J=6.6, 2.6 Hz, 1H), 7.74-7.71 (m, 1H), 7.67 (d, J=7.2 Hz, 1H), 7.60 (s, 1H), 7.52 (t, J=7.6 Hz, 1H), 7.36 (t, J=8.9 Hz, 1H), 7.28 (s, 1H), 5.15 (s, 1H), 5.03 (s, 1H), 4.45 (s, 1H), 4.41 (d, J=17.6 Hz, 1H), 4.10 (s, 3H), 3.95 (s, 2H), 3.82-3.33 (m, 6H), 3.29 (s, 2H), 3.17-3.04 (m, 2H), 2.94-2.85 (m, 1H), 2.78-2.74 (m, 1H), 2.53 (dt, J=17.9, 10.9 Hz, 1H), 2.31 (s, 2H), 2.18-2.13 (m, 1H), 1.69 (s, 4H). HRMS (ESI) m/z: calcd for C39H42ClFN7O6S+ [M+H]+, 790.2584; found, 790.2581.
Referring to the method of example 1, the target compound (SIAIS293050) was prepared by using Gefitinib derivative C and intermediate LM (SIAIS171134). (yellow solid, 8.7 mg, yield 48%). 1H NMR (500 MHz, MeOD) δ 8.73 (s, 1H), 8.35 (s, 1H), 8.00 (dd, J=6.6, 2.4 Hz, 1H), 7.74 (s, 1H), 7.65-7.59 (m, 2H), 7.54-7.48 (m, 1H), 7.36 (t, J=8.9 Hz, 1H), 7.27 (s, 1H), 5.16 (dd, J=13.4, 5.1 Hz, 1H), 5.08 (s, 1H), 4.45 (d, J=17.3 Hz, 1H), 4.38 (d, J=17.3 Hz, 1H), 4.10 (s, 3H), 4.00 (s, 2H), 3.60 (dd, J=43.0, 36.5 Hz, 4H), 3.31-3.22 (m, 4H), 3.12-3.01 (m, 2H), 2.91 (ddd, J=17.4, 14.3, 7.5 Hz, 1H), 2.84-2.75 (m, 1H), 2.56-2.46 (m, 1H), 2.33 (s, 2H), 2.21-2.14 (m, 1H), 1.69-1.60 (m, 2H), 1.52 (dt, J=18.4, 5.4 Hz, 4H), 1.37 (dt, J=15.7, 8.0 Hz, 2H). HRMS (ESI) m/z: calcd for C41H46ClFN7O6S+ [M+H]+, 818.2897; found, 818.2892.
Referring to the method of example 1, the target compound (SIAIS293051) was prepared by using Gefitinib derivative C and intermediate LM (SIAIS171135). (yellow solid, 8.5 mg, yield 46%). 1H NMR (500 MHz, MeOD) δ 8.72 (s, 1H), 8.31-8.21 (m, 1H), 7.99 (s, 1H), 7.72 (s, 1H), 7.61 (d, J=7.6 Hz, 2H), 7.53-7.48 (m, 1H), 7.36 (t, J=8.9 Hz, 1H), 7.26 (s, 1H), 5.16 (dd, J=13.3, 5.2 Hz, 1H), 5.03 (s, 1H), 4.44 (d, J=17.3 Hz, 1H), 4.38 (d, J=17.2 Hz, 1H), 4.09 (s, 3H), 4.00 (s, 2H), 3.46 (dd, J=10.8, 9.1 Hz, 6H), 3.25 (t, J=7.1 Hz, 2H), 3.08-3.03 (m, 2H), 2.90 (m, 1H), 2.82-2.76 (m, 1H), 2.55-2.48 (m, 1H), 2.34 (s, 2H), 2.19 (t, J=7.5 Hz, 1H), 1.64 (dd, J=14.8, 7.2 Hz, 2H), 1.52 (d, J=6.6 Hz, 2H), 1.46 (s, 2H), 1.33 (s, 4H). HRMS (ESI) m/z: calcd for C42H48ClFN7O6S+ [M+H]+, 832.3054; found, 832.3052.
Referring to the method of example 1, the target compound (SIAIS293052) was prepared by using Gefitinib derivative C and intermediate LM (SIAIS171136). (yellow solid, 9.1 mg, yield 49%). 1H NMR (500 MHz, MeOD) δ 8.72 (s, 1H), 8.35 (s, 1H), 8.00 (dd, J=6.6, 2.6 Hz, 1H), 7.73 (s, 1H), 7.64-7.60 (m, 2H), 7.53-7.48 (m, 1H), 7.36 (t, J=8.9 Hz, 1H), 7.27 (s, 1H), 5.16 (dd, J=13.3, 5.2 Hz, 1H), 5.08 (s, 1H), 4.44 (d, J=17.3 Hz, 1H), 4.38 (d, J=17.3 Hz, 1H), 4.10 (s, 3H), 4.01 (s, 2H), 3.95-3.32 (m, 6H), 3.24 (dd, J=13.0, 5.9 Hz, 2H), 3.11-3.01 (m, 2H), 2.94-2.86 (m, 1H), 2.78 (m, 1H), 2.52 (dt, J=13.4, 8.6 Hz, 1H), 2.34 (s, 2H), 2.20-2.16 (m, 1H), 1.70-1.63 (m, 2H), 1.56-1.44 (m, 4H), 1.39-1.31 (m, 6H). HRMS (ESI) m/z: calcd for C43H50ClFN7O6S+ [M+H]+, 846.3210; found, 846.3217.
Referring to the method of example 1, the target compound (SIAIS293067) was prepared by using Sapitinib derivative A and intermediate LM (SIAIS171131). (yellow solid, 9.1 mg, yield 49%). 1H NMR (500 MHz, MeOD) δ 8.70 (d, J=5.0 Hz, 1H), 8.40-8.26 (m, 1H), 7.69-7.62 (m, 2H), 7.53 (s, 3H), 7.31 (d, J=13.9 Hz, 2H), 5.14 (s, 1H), 4.42 (dd, J=30.2, 16.1 Hz, 2H), 4.11 (d, J=14.3 Hz, 3H), 3.97 (d, J=7.6 Hz, 2H), 3.67-3.36 (m, 5H), 3.09 (s, 2H), 2.95-2.84 (m, 1H), 2.78 (d, J=16.3 Hz, 1H), 2.58-2.43 (m, 2H), 2.33 (s, 2H), 2.18 (s, 1H), 2.09 (s, 1H), 1.70 (s, 4H). HRMS (ESI) m/z: calcd for C39H42ClFN7O6S+ [M+H]+, 790.2584; found, 790.2582.
Referring to the method of example 1, the target compound (SIAIS293068) was prepared by using Sapitinib derivative A and intermediate LM (SIAIS171134). (yellow solid, 9.1 mg, yield 49%). 1H NMR (500 MHz, MeOD) δ 8.70 (s, 1H), 8.30 (d, J=51.9 Hz, 1H), 7.62 (d, J=7.5 Hz, 2H), 7.55 (d, J=7.2 Hz, 3H), 7.31 (s, 2H), 5.15 (s, 1H), 4.45 (d, J=18.1 Hz, 1H), 4.39 (d, J=17.6 Hz, 1H), 4.11 (s, 3H), 4.00 (s, 2H), 3.81-3.42 (m, 5H), 3.05 (s, 2H), 2.90 (s, 1H), 2.78 (d, J=18.1 Hz, 1H), 2.52 (s, 1H), 2.34 (s, 2H), 2.22-1.90 (m, 3H), 1.75-1.71 (m, 2H), 1.50 (s, 4H), 1.36-1.32 (m, 4H). HRMS (ESI) m/z: calcd for C41H46ClFN7O6S+ [M+H]+, 818.2897; found, 818.2895.
Referring to the method of example 1, the target compound (SIAIS293069) was prepared by using Sapitinib derivative A and intermediate LM (SIAIS171135). (yellow solid, 9.1 mg, yield 49%). 1H NMR (500 MHz, MeOD) δ 8.69 (s, 1H), 8.23 (s, 1H), 7.63 (d, J=7.6 Hz, 2H), 7.53 (dq, J=14.8, 7.4 Hz, 3H), 7.32-7.27 (m, 2H), 5.17 (dd, J=13.2, 5.0 Hz, 1H), 4.45 (d, J=17.4 Hz, 1H), 4.39 (d, J=17.2 Hz, 1H), 4.11 (s, 3H), 4.00 (s, 2H), 3.48 (dd, J=57.2, 32.1 Hz, 5H), 3.24 (t, J=7.0 Hz, 2H), 3.10-3.01 (m, 2H), 2.94-2.87 (m, 1H), 2.79 (d, J=16.0 Hz, 1H), 2.60-2.45 (m, 2H), 2.36 (s, 2H), 2.20-2.07 (m, 2H), 1.69-1.62 (m, 2H), 1.50 (dd, J=18.8, 11.8 Hz, 4H), 1.35 (s, 4H). HRMS (ESI) m/z: calcd for C42H48ClFN7O6S+ [M+H]+, 832.3054; found, 832.3044.
Referring to the method of example 1, the target compound (SIAIS293070) was prepared by using Sapitinib derivative A and intermediate LM (SIAIS171136). (yellow solid, 9.1 mg, yield 49%). 1H NMR (500 MHz, MeOD) δ 8.67 (s, 1H), 8.21 (s, 1H), 7.62 (d, J=7.6 Hz, 2H), 7.53-7.48 (m, 3H), 7.33-7.27 (m, 2H), 5.18 (dd, J=13.2, 5.0 Hz, 1H), 4.46 (d, J=17.4 Hz, 1H), 4.36 (d, J=17.2 Hz, 1H), 4.10 (s, 3H), 4.00 (s, 2H), 3.47-3.43 (m, 5H), 3.23 (t, J=7.0 Hz, 2H), 3.10-3.01 (m, 2H), 2.93-2.87 (m, 1H), 2.77 (d, J=16.0 Hz, 1H), 2.60-2.46 (m, 2H), 2.34 (s, 2H), 2.21-2.07 (m, 2H), 1.68-1.62 (m, 2H), 1.50-1.44 (m, 4H), 1.35 (s, 4H). HRMS (ESI) m/z: calcd for C43H50ClFN7O6S+ [M+H]+, 846.3210; found, 846.3212.
Referring to the method of example 1, the target compound (SIAIS337052) was prepared by using Osimertinib derivative SIAIS337051 and intermediate LM (SIAIS171090). (yellow solid, 9.2 mg, yield 45%). 1H NMR (500 MHz, DMSO-d6) δ 10.98 (d, J=7.0 Hz, 1H), 9.28 (s, 1H), 8.82 (s, 1H), 8.22 (s, 2H), 7.66 (d, J=7.7 Hz, 1H), 7.58 (t, J=9.4 Hz, 2H), 7.50 (dt, J=15.4, 7.7 Hz, 1H), 7.44-7.38 (m, 1H), 7.30 (s, 1H), 7.19 (s, 1H), 7.05 (d, J=38.4 Hz, 1H), 6.69 (dt, J=19.8, 9.5 Hz, 1H), 6.19 (d, J=17.1 Hz, 1H), 5.69 (d, J=10.3 Hz, 1H), 5.12 (dd, J=13.4, 5.2 Hz, 1H), 4.39 (d, J=17.5 Hz, 1H), 4.25 (d, J=17.3 Hz, 1H), 4.11 (s, 2H), 3.93 (s, 3H), 3.82 (s, 3H), 3.50 (d, J=7.1 Hz, 4H), 3.03 (s, 2H), 2.90 (t, J=13.8 Hz, 1H), 2.84-2.74 (m, 4H), 2.59 (s, 1H), 2.43 (s, 1H), 2.02-1.96 (m, 1H). HRMS (ESI) m/z: calcd for C42H44N9O6S+ [M+H]+, 802.3130; found, 802.3132.
Referring to the method of example 1, the target compound (SIAIS337053) was prepared by using Osimertinib derivative SIAIS337051 and intermediate LM (SIAIS171086). (yellow solid, 9.7 mg, yield 44%). 1H NMR (500 MHz, DMSO-d6) δ 10.98 (s, 1H), 9.25 (s, 1H), 8.82 (s, 1H), 8.22 (s, 2H), 7.57 (ddt, J=26.7, 15.6, 8.4 Hz, 4H), 7.41 (d, J=6.7 Hz, 1H), 7.29 (d, J=8.6 Hz, 1H), 7.19 (s, 1H), 7.02 (d, J=22.5 Hz, 1H), 6.70 (dd, J=16.9, 9.9 Hz, 1H), 6.19 (d, J=17.0 Hz, 1H), 5.71 (d, J=10.8 Hz, 1H), 5.11 (dd, J=13.4, 6.0 Hz, 1H), 4.32 (d, J=17.1 Hz, 1H), 4.19 (d, J=8.6 Hz, 1H), 3.93 (s, 3H), 3.84 (s, 3H), 3.52-3.38 (m, 4H), 3.23 (d, J=8.0 Hz, 2H), 3.13 (s, 1H), 2.86 (s, 3H), 2.80 (d, J=6.5 Hz, 3H), 2.71-2.61 (m, 3H), 2.40 (s, 1H), 1.98 (s, 1H). HRMS (ESI) m/z: calcd for C43H46N9O6S+ [M+H]+, 816.3286; found, 816.3277.
Referring to the method of example 1, the target compound (SIAIS337054) was prepared by using Osimertinib derivative SIAIS337051 and intermediate LM (SIAIS171089). (yellow solid, 9.5 mg, yield 51%). 1H NMR (500 MHz, DMSO-d6) δ 10.98 (s, 1H), 9.32 (s, 1H), 8.81 (s, 1H), 8.22 (s, 2H), 7.65 (d, J=7.5 Hz, 1H), 7.58-7.52 (m, 3H), 7.40 (t, J=6.2 Hz, 1H), 7.29 (d, J=8.2 Hz, 1H), 7.18 (s, 1H), 7.07 (s, 1H), 6.75-6.63 (m, 1H), 6.24-6.15 (m, 1H), 5.70 (d, J=10.3 Hz, 1H), 5.11 (dd, J=12.9, 6.3 Hz, 1H), 4.34 (d, J=17.3 Hz, 1H), 4.20 (d, J=17.3 Hz, 1H), 3.93 (s, 3H), 3.83 (s, 3H), 3.49 (d, J=6.9 Hz, 4H), 3.16-3.09 (m, 2H), 3.00 (s, 1H), 2.89 (s, 3H), 2.80 (s, 3H), 2.43-2.33 (m, 3H), 2.01-1.98 (m, 1H), 1.84-1.72 (m, 2H). HRMS (ESI) m/z: calcd for C44H48N9O6S+ [M+H]+, 830.3443; found, 830.3441.
Referring to the method of example 1, the target compound (SIAIS337055) was prepared by using Osimertinib derivative SIAIS337051 and intermediate LM (SIAIS171079). (yellow solid, 9.9 mg, yield 48%). 1H NMR (500 MHz, DMSO-d6) δ 10.99 (s, 1H), 9.28 (s, 1H), 8.82 (s, 1H), 8.23 (s, 2H), 7.64 (d, J=7.5 Hz, 1H), 7.58-7.53 (m, 3H), 7.41 (t, J=6.2 Hz, 1H), 7.28 (d, J=8.2 Hz, 1H), 7.18 (s, 1H), 7.08 (s, 1H), 6.76-6.63 (m, 1H), 6.25-6.15 (m, 1H), 5.71 (d, J=10.3 Hz, 1H), 5.13 (dd, J=12.9, 6.3 Hz, 1H), 4.36 (d, J=17.3 Hz, 1H), 4.22 (d, J=17.3 Hz, 1H), 3.95 (s, 3H), 3.84 (s, 3H), 3.51-3.47 (m, 4H), 3.17-3.09 (m, 2H), 3.01 (s, 1H), 2.89 (s, 3H), 2.82 (s, 3H), 2.44-2.33 (m, 3H), 2.02-1.99 (m, 1H), 1.86-1.72 (m, 4H). HRMS (ESI) m/z: calcd for C45H50N9O6S+ [M+H]+, 844.3599; found, 844.3601.
Referring to the method of example 1, the target compound (SIAIS337056) was prepared by using Osimertinib derivative SIAIS337051 and intermediate LM (SIAIS171091). (yellow solid, 9.3 mg, yield 43%). 1H NMR (500 MHz, DMSO-d6) δ 10.97 (s, 1H), 9.25 (s, 1H), 8.84 (s, 1H), 8.25 (s, 2H), 7.66 (d, J=7.5 Hz, 1H), 7.56-7.53 (m, 3H), 7.42 (t, J=6.2 Hz, 1H), 7.29 (d, J=8.2 Hz, 1H), 7.16 (s, 1H), 7.05 (s, 1H), 6.77-6.63 (m, 1H), 6.24-6.15 (m, 1H), 5.72 (d, J=10.3 Hz, 1H), 5.14 (dd, J=12.9, 6.3 Hz, 1H), 4.35 (d, J=17.3 Hz, 1H), 4.24 (d, J=17.3 Hz, 1H), 3.97 (s, 3H), 3.85 (s, 3H), 3.51-3.46 (m, 4H), 3.18-3.11 (m, 2H), 3.04 (s, 1H), 2.88 (s, 3H), 2.83 (s, 3H), 2.45-2.33 (m, 3H), 2.03-1.99 (m, 1H), 1.88-1.71 (m, 6H). HRMS (ESI) m/z: calcd for C46H52N9O6S+ [M+H]+, 858.3756; found, 858.3759.
Referring to the method of example 1, the target compound (SIAIS337057) was prepared by using Osimertinib derivative SIAIS337051 and intermediate LM (SIAIS171092). (yellow solid, 8.8 mg, yield 43%). 1H NMR (500 MHz, DMSO-d6) δ 10.99 (s, 1H), 9.26 (s, 1H), 8.84 (s, 1H), 8.26 (s, 2H), 7.68 (d, J=7.5 Hz, 1H), 7.57-7.53 (m, 3H), 7.43 (t, J=6.2 Hz, 1H), 7.28 (d, J=8.2 Hz, 1H), 7.15 (s, 1H), 7.07 (s, 1H), 6.79-6.63 (m, 1H), 6.26-6.15 (m, 1H), 5.71 (d, J=10.3 Hz, 1H), 5.14 (dd, J=12.9, 6.3 Hz, 1H), 4.36 (d, J=17.3 Hz, 1H), 4.25 (d, J=17.3 Hz, 1H), 3.98 (s, 3H), 3.86 (s, 3H), 3.52-3.46 (m, 4H), 3.19-3.11 (m, 2H), 3.05 (s, 1H), 2.89 (s, 3H), 2.84 (s, 3H), 2.46-2.33 (m, 3H), 2.04-1.98 (m, 1H), 1.89-1.71 (m, 8H). HRMS (ESI) m/z: calcd for C47H54N9O6S+ [M+H]+, 872.3912; found, 872.3904.
Referring to the method of example 1, the target compound (SIAIS337059) was prepared by using Osimertinib derivative SIAIS337051 and intermediate LM (SIAIS299138). (yellow solid, 7.9 mg, yield 38%). 1H NMR (500 MHz, DMSO-d6) δ 10.97 (s, 1H), 9.25 (s, 1H), 8.83 (s, 1H), 8.25 (s, 2H), 7.67 (d, J=7.5 Hz, 1H), 7.58-7.53 (m, 3H), 7.42-7.40 (m, 1H), 7.26 (d, J=8.2 Hz, 1H), 7.14 (s, 1H), 7.09 (s, 1H), 6.77-6.63 (m, 1H), 6.28-6.15 (m, 1H), 5.72 (d, J=10.3 Hz, 1H), 5.16 (dd, J=12.9, 6.3 Hz, 1H), 4.33 (d, J=17.3 Hz, 1H), 4.23 (d, J=17.3 Hz, 1H), 3.97 (s, 3H), 3.84 (s, 3H), 3.55-3.46 (m, 4H), 3.18-3.11 (m, 2H), 3.06 (s, 1H), 2.87 (s, 3H), 2.83 (s, 3H), 2.48-2.33 (m, 3H), 2.02-1.98 (m, 1H), 1.86-1.70 (m, 10H). HRMS (ESI) m/z: calcd for C49H58N9O6S+ [M+H]+, 900.4225; found, 900.4217.
Referring to the method of example 1, the target compound (SIAIS337060) was prepared by using Osimertinib derivative SIAIS337051 and intermediate LM (SIAIS299135). (yellow solid, 9.5 mg, yield 42%). 1H NMR (500 MHz, DMSO-d6) δ 10.99 (s, 1H), 9.26 (s, 1H), 8.85 (s, 1H), 8.27 (s, 2H), 7.68 (d, J=7.5 Hz, 1H), 7.59-7.53 (m, 3H), 7.46-7.40 (m, 1H), 7.25 (d, J=8.2 Hz, 1H), 7.13 (s, 1H), 7.08 (s, 1H), 6.79-6.63 (m, 1H), 6.26-6.15 (m, 1H), 5.71 (d, J=10.3 Hz, 1H), 5.13 (dd, J=12.9, 6.3 Hz, 1H), 4.34 (d, J=17.3 Hz, 1H), 4.25 (d, J=17.3 Hz, 1H), 3.98 (s, 3H), 3.83 (s, 3H), 3.57-3.46 (m, 4H), 3.19-3.13 (m, 2H), 3.08 (s, 1H), 2.88 (s, 3H), 2.85 (s, 3H), 2.49-2.33 (m, 3H), 2.03-1.99 (m, 1H), 1.87-1.70 (m, 12H). HRMS (ESI) m/z: calcd for C50H60N9O6S+ [M+H]+, 914.4382; found, 914.4386.
Referring to the method of example 1, the target compound (SIAIS337061) was prepared by using Osimertinib derivative SIAIS337051 and intermediate LM (SIAIS122009). (yellow solid, 9.7 mg, yield 43%). 1H NMR (500 MHz, DMSO-d6) δ 10.98 (s, 1H), 9.25 (s, 1H), 8.84 (s, 1H), 8.27 (s, 2H), 7.69 (d, J=7.5 Hz, 1H), 7.58-7.53 (m, 3H), 7.48-7.40 (m, 1H), 7.27 (d, J=8.2 Hz, 1H), 7.14 (s, 1H), 7.09 (s, 1H), 6.79-6.65 (m, 1H), 6.28-6.15 (m, 1H), 5.72 (d, J=10.3 Hz, 1H), 5.15 (dd, J=12.9, 6.3 Hz, 1H), 4.35 (d, J=17.3 Hz, 1H), 4.26 (d, J=17.3 Hz, 1H), 3.99 (s, 3H), 3.85 (s, 3H), 3.59-3.46 (m, 4H), 3.19-3.12 (m, 2H), 3.08 (s, 1H), 2.89 (s, 3H), 2.87 (s, 3H), 2.49-2.36 (m, 3H), 2.04-2.00 (m, 1H), 1.89-1.70 (m, 14H). HRMS (ESI) m/z: calcd for C51H62N9O6S+ [M+H]+, 928.4538; found, 928.4544.
Referring to the method of example 1, the target compound (SIAIS337074) was prepared by using Osimertinib derivative SIAIS337051 and intermediate LM (SIAIS1213129). (yellow solid, 9.8 mg, yield 44%). 1H NMR (500 MHz, Methanol-d4) δ 8.58 (s, 1H), 8.13-8.01 (m, 2H), 7.56-7.25 (m, 8H), 7.15 (s, 1H), 6.61 (d, J=11.8 Hz, 1H), 6.49 (d, J=16.8 Hz, 1H), 5.93 (d, J=9.7 Hz, 1H), 5.17 (dd, J=13.2, 5.0 Hz, 1H), 4.39-4.26 (m, 4H), 3.98 (d, J=8.6 Hz, 3H), 3.89 (s, 3H), 3.75-3.55 (m, 8H), 3.07 (d, J=19.0 Hz, 6H), 2.87-2.79 (m, 1H), 2.74-2.70 (m, 1H), 2.43-2.39 (m, 1H), 2.09 (s, 1H). HRMS (ESI) m/z: calcd for C44H48N9O7S+ [M+H]+, 846.3392; found, 846.3395.
Referring to the method of example 1, the target compound (SIAIS337075) was prepared by using Osimertinib derivative SIAIS337051 and intermediate LM (SIAIS1213131). (yellow solid, 8.8 mg, yield 41%). 1H NMR (500 MHz, Methanol-d4) δ 8.59 (s, 1H), 8.14-8.01 (m, 2H), 7.58-7.25 (m, 8H), 7.17 (s, 1H), 6.60 (d, J=11.8 Hz, 1H), 6.48 (d, J=16.8 Hz, 1H), 5.92 (d, J=9.7 Hz, 1H), 5.16 (dd, J=13.2, 5.0 Hz, 1H), 4.37-4.25 (m, 4H), 3.97 (d, J=8.6 Hz, 3H), 3.88 (s, 3H), 3.74-3.52 (m, 12H), 3.05 (d, J=19.0 Hz, 6H), 2.86-2.78 (m, 1H), 2.73-2.70 (m, 1H), 2.42-2.39 (m, 1H), 2.07 (s, 1H). HRMS (ESI) m/z: calcd for C46H52N9O8S+ [M+H]+, 890.3654; found, 890.3658.
Referring to the method of example 1, the target compound (SIAIS337076) was prepared by using Osimertinib derivative SIAIS337051 and intermediate LM (SIAIS1213133). (yellow solid, 10.1 mg, yield 39%). 1H NMR (500 MHz, Methanol-d4) δ 8.60 (s, 1H), 8.15-8.02 (m, 2H), 7.59-7.25 (m, 8H), 7.19 (s, 1H), 6.61 (d, J=11.8 Hz, 1H), 6.50 (d, J=16.8 Hz, 1H), 5.94 (d, J=9.7 Hz, 1H), 5.18 (dd, J=13.2, 5.0 Hz, 1H), 4.38-4.22 (m, 4H), 3.98 (d, J=8.6 Hz, 3H), 3.92 (s, 3H), 3.76-3.52 (m, 16H), 3.07 (d, J=19.0 Hz, 6H), 2.87-2.79 (m, 1H), 2.71 (d, J=17.7 Hz, 1H), 2.43-2.40 (m, 1H), 2.08 (s, 1H). HRMS (ESI) m/z: calcd for C48H56N9O9S+ [M+H]+, 934.3916; found, 934.3919.
Referring to the method of example 1, the target compound (SIAIS337077) was prepared by using Osimertinib derivative SIAIS337051 and intermediate LM (SIAIS1213135). (yellow solid, 10.9 mg, yield 43%). 1H NMR (500 MHz, Methanol-d4) δ 8.62 (s, 1H), 8.17-8.02 (m, 2H), 7.61-7.26 (m, 8H), 7.21 (s, 1H), 6.63 (d, J=11.8 Hz, 1H), 6.51 (d, J=16.8 Hz, 1H), 5.96 (d, J=9.7 Hz, 1H), 5.17 (dd, J=13.2, 5.0 Hz, 1H), 4.39-4.24 (m, 4H), 3.99 (d, J=8.6 Hz, 3H), 3.94 (s, 3H), 3.78-3.52 (m, 20H), 3.09-3.05 (m, 6H), 2.88-2.79 (m, 1H), 2.72 (d, J=17.7 Hz, 1H), 2.45-2.41 (m, 1H), 2.09 (s, 1H). HRMS (ESI) m/z: calcd for C50H60N9O10S+ [M+H]+, 978.4178; found, 978.4172.
Referring to the method of example 1, the target compound (SIAIS337078) was prepared by using Osimertinib derivative SIAIS337051 and intermediate LM (SIAIS1213137). (yellow solid, 9.9 mg, yield 34%). 1H NMR (500 MHz, Methanol-d4) δ 8.61 (s, 1H), 8.16-8.02 (m, 2H), 7.60-7.31 (m, 8H), 7.22 (s, 1H), 6.62 (d, J=11.8 Hz, 1H), 6.50 (d, J=16.8 Hz, 1H), 5.95 (d, J=9.7 Hz, 1H), 5.16 (dd, J=13.2, 5.0 Hz, 1H), 4.38-4.27 (m, 4H), 3.98 (d, J=8.6 Hz, 3H), 3.93 (s, 3H), 3.77-3.52 (m, 24H), 3.08-3.05 (m, 6H), 2.87-2.79 (m, 1H), 2.73 (d, J=17.7 Hz, 1H), 2.45-2.41 (m, 1H), 2.08 (s, 1H). HRMS (ESI) m/z: calcd for C52H64N9O11S+ [M+H]+, 1022.4441; found, 1022.4437.
Referring to the method of example 1, the target compound (SIAIS337079) was prepared by using Osimertinib derivative SIAIS337051 and intermediate LM (SIAIS213132). (yellow solid, 9.8 mg, yield 46%). 1H NMR (500 MHz, Methanol-d4) δ 8.52 (s, 1H), 8.26 (s, 1H), 8.12-7.95 (m, 2H), 7.50 (d, J=34.1 Hz, 2H), 7.41 (d, J=6.5 Hz, 1H), 7.37-7.25 (m, 4H), 7.05 (d, J=10.1 Hz, 1H), 6.55 (d, J=10.3 Hz, 1H), 6.48 (d, J=10.5 Hz, 1H), 5.85 (d, J=10.0 Hz, 1H), 5.13 (dd, J=13.2, 5.0 Hz, 1H), 4.47 (d, J=17.4 Hz, 1H), 4.35 (d, J=17.2 Hz, 1H), 4.01 (s, 3H), 3.97 (s, 3H), 3.55-3.49 (m, 4H), 3.23-3.21 (m, 2H), 2.95 (s, 3H), 2.87-2.85 (m, 3H), 2.83 (s, 3H), 2.75-2.69 (m, 1H), 2.31-2.19 (m, 1H), 2.07-1.93 (m, 3H). HRMS (ESI) m/z: calcd for C43H48N9O5S+ [M+H]+, 802.3494; found, 802.3493.
Referring to the method of example 1, the target compound (SIAIS337081) was prepared by using Osimertinib derivative SIAIS337051 and intermediate LM (SIAIS213135). (yellow solid, 9.7 mg, yield 43%). 1H NMR (500 MHz, Methanol-d4) δ 8.42 (d, J=11.4 Hz, 1H), 8.28 (s, 1H), 7.96 (d, J=39.8 Hz, 2H), 7.64 (d, J=7.5 Hz, 1H), 7.52 (s, 2H), 7.27 (s, 3H), 7.11 (d, J=34.9 Hz, 2H), 6.52 (s, 2H), 5.87 (s, 1H), 5.18 (dd, J=13.2, 5.0 Hz, 1H), 4.46 (d, J=17.4 Hz, 1H), 4.36 (d, J=17.2 Hz, 1H), 4.08 (s, 3H), 3.98 (s, 3H), 3.55-3.49 (m, 4H), 3.28-3.23 (m, 2H), 2.95 (s, 3H), 2.86-2.82 (m, 3H), 2.80 (s, 3H), 2.75-2.69 (m, 1H), 2.36-2.17 (m, 1H), 2.09-1.95 (m, 7H). HRMS (ESI) m/z: calcd for C45H52N9O5S+ [M+H]+, 830.3807; found, 830.3803.
Referring to the method of example 1, the target compound (SIAIS337082) was prepared by using Osimertinib derivative SIAIS337051 and intermediate LM (SIAIS1216133). (yellow solid, 9.9 mg, yield 47%). 1H NMR (500 MHz, Methanol-d4) δ 8.47 (d, J=13.6 Hz, 1H), 8.31 (s, 1H), 8.05-8.01 (m, 2H), 7.54 (d, J=11.4 Hz, 2H), 7.35-7.31 (m, 5H), 7.07 (s, 1H), 6.58 (s, 1H), 6.48 (d, J=17.2 Hz, 1H), 5.85 (s, 1H), 5.17 (dd, J=13.2, 5.0 Hz, 1H), 4.45 (d, J=17.4 Hz, 1H), 4.35 (d, J=17.2 Hz, 1H), 4.01 (s, 3H), 3.96 (s, 3H), 3.53-3.46 (m, 4H), 3.26-3.21 (m, 2H), 2.93 (s, 3H), 2.85-2.82 (m, 3H), 2.79 (s, 3H), 2.74-2.62 (m, 1H), 2.34-2.15 (m, 1H), 2.06-1.93 (m, 9H). HRMS (ESI) m/z: calcd for C46H54N9O5S+ [M+H]+, 844.3963; found, 844.3967.
Referring to the method of example 1, the target compound (SIAIS337083) was prepared by using Osimertinib derivative SIAIS337051 and intermediate LM (SIAIS1216135). (yellow solid, 8.7 mg, yield 35%). 1H NMR (500 MHz, Methanol-d4) δ 8.62 (d, J=14.0 Hz, 1H), 8.48 (s, 1H), 8.24 (s, 1H), 8.14 (t, J=7.7 Hz, 1H), 7.73-7.64 (m, 2H), 7.56-7.40 (m, 4H), 7.23 (s, 1H), 6.83-6.71 (m, 1H), 6.65 (d, J=16.9 Hz, 1H), 6.02 (d, J=10.4 Hz, 1H), 5.22 (dd, J=13.2, 5.0 Hz, 1H), 4.48 (d, J=17.4 Hz, 1H), 4.37 (d, J=17.2 Hz, 1H), 4.09 (s, 3H), 3.99 (s, 3H), 3.59-3.48 (m, 4H), 3.29-3.25 (m, 2H), 2.98 (s, 3H), 2.87-2.85 (m, 3H), 2.83 (s, 3H), 2.79-2.68 (m, 1H), 2.37-2.15 (m, 1H), 2.09-1.96 (m, 11H). HRMS (ESI) m/z: calcd for C47H56N9O5S+ [M+H]+, 858.4120; found, 858.4122.
Referring to the method of example 1, the target compound (SIAIS337084) was prepared by using Osimertinib derivative SIAIS337051 and intermediate LM (SIAIS1216137). (yellow solid, 9.4 mg, yield 43%). 1H NMR (500 MHz, Methanol-d4) δ 8.52 (s, 1H), 8.34 (s, 1H), 8.03 (s, 1H), 7.98 (d, J=2.1 Hz, 1H), 7.61 (d, J=7.0 Hz, 1H), 7.54 (d, J=8.4 Hz, 1H), 7.45 (d, J=9.4 Hz, 1H), 7.42 (dd, J=8.2, 1.9 Hz, 2H), 7.37 (d, J=6.2 Hz, 1H), 7.34-7.26 (m, 2H), 7.08 (s, 1H), 6.66-6.57 (m, 1H), 6.49 (d, J=16.9 Hz, 1H), 5.87 (d, J=10.0 Hz, 1H), 5.16 (dd, J=13.2, 5.0 Hz, 1H), 4.42 (d, J=17.4 Hz, 1H), 4.33 (d, J=17.2 Hz, 1H), 4.05 (s, 3H), 3.95 (s, 3H), 3.56-3.46 (m, 4H), 3.25-3.25 (m, 2H), 2.94 (s, 3H), 2.87-2.85 (m, 3H), 2.81 (s, 3H), 2.77-2.68 (m, 1H), 2.35-2.17 (m, 1H), 2.07-1.96 (m, 13H). HRMS (ESI) m/z: calcd for C48H58N9O5S+ [M+H]+, 872.4276; found, 872.4278.
Referring to the method of example 1, the target compound (SIAIS337085) was prepared by using Osimertinib derivative SIAIS337051 and intermediate LM (SIAIS1220059). (yellow solid, 9.3 mg, yield 46%). 1H NMR (500 MHz, Methanol-d4) δ 8.51 (d, J=15.2 Hz, 1H), 8.35 (s, 1H), 8.04 (s, 2H), 7.56 (d, J=27.2 Hz, 3H), 7.45-7.25 (m, 5H), 7.09 (s, 1H), 6.61 (d, J=12.3 Hz, 1H), 6.50 (d, J=16.8 Hz, 1H), 5.88 (d, J=10.3 Hz, 1H), 5.16 (dd, J=13.2, 5.0 Hz, 1H), 4.45 (d, J=17.4 Hz, 1H), 4.34 (d, J=17.2 Hz, 1H), 4.03 (s, 3H), 3.93 (s, 3H), 3.57-3.46 (m, 4H), 3.27-3.28 (m, 2H), 2.93 (s, 3H), 2.86-2.82 (m, 3H), 2.80 (s, 3H), 2.75-2.68 (m, 1H), 2.34-2.16 (m, 1H), 2.08-1.96 (m, 15H). HRMS (ESI) m/z: calcd for C49H60N9O5S+ [M+H]+, 886.4433; found, 886.4431.
Referring to the method of example 1, the target compound (SIAIS337086) was prepared by using Osimertinib derivative SIAIS337051 and intermediate LM (SIAIS1220013). (yellow solid, 9.8 mg, yield 47%). 1H NMR (500 MHz, MeOD) δ 8.53 (d, J=15.2 Hz, 1H), 8.37 (s, 1H), 8.05 (s, 2H), 7.57 (d, J=27.2 Hz, 3H), 7.46-7.25 (m, 5H), 7.08 (s, 1H), 6.60 (d, J=12.3 Hz, 1H), 6.48 (d, J=16.8 Hz, 1H), 5.87 (d, J=10.3 Hz, 1H), 5.13 (dd, J=13.2, 5.0 Hz, 1H), 4.42 (d, J=17.4 Hz, 1H), 4.33 (d, J=17.2 Hz, 1H), 4.01 (s, 3H), 3.92 (s, 3H), 3.56-3.43 (m, 4H), 3.25-3.21 (m, 2H), 2.91 (s, 3H), 2.85-2.82 (m, 3H), 2.80 (s, 3H), 2.74-2.68 (m, 1H), 2.32-2.16 (m, 1H), 2.08-1.96 (m, 17H). HRMS (ESI) m/z: calcd for C50H62N9O5S+ [M+H]+, 900.4589; found, 900.4586.
Referring to the method of example 1, the target compound (SIAIS337087) was prepared by using Osimertinib derivative SIAIS337051 and intermediate LM (SIAIS1220015). (yellow solid, 8.7 mg, yield 42%). 1H NMR (500 MHz, Methanol-d4) δ 8.53 (s, 1H), 8.33 (s, 1H), 8.03 (s, 2H), 7.60 (d, J=7.4 Hz, 1H), 7.52 (d, J=7.3 Hz, 2H), 7.46 (t, J=7.8 Hz, 1H), 7.39 (d, J=6.9 Hz, 1H), 7.28 (dt, J=27.3, 7.4 Hz, 2H), 7.06 (s, 1H), 6.59 (dd, J=16.9, 10.0 Hz, 1H), 6.48 (d, J=17.0 Hz, 1H), 5.85 (d, J=10.2 Hz, 1H), 5.17-5.12 (m, 1H), 4.37 (t, J=5.6 Hz, 2H), 4.00 (s, 3H), 3.94 (s, 3H), 3.57-3.42 (m, 3H), 3.11 (q, J=7.3 Hz, 3H), 2.92 (dd, J=17.1, 4.2 Hz, 5H), 2.81 (d, J=1.5 Hz, 3H), 2.76 (d, J=17.3 Hz, 1H), 2.49 (t, J=6.7 Hz, 1H), 2.20-2.11 (m, 1H), 2.03 (d, J=6.7 Hz, 1H), 1.55 (s, 4H), 1.30 (s, 4H), 1.08 (d, J=35.9 Hz, 10H). HRMS (ESI) m/z: calcd for C51H64N9O5S+ [M+H]+, 914.4746; found, 914.4743.
Referring to the method of example 1, the target compound (SIAIS337088) was prepared by using Osimertinib derivative SIAIS337051 and intermediate LM (SIAIS1220141). (yellow solid, 9.8 mg, yield 43%). 1H NMR (500 MHz, Methanol-d4) δ 8.58 (s, 1H), 8.57-8.47 (m, 1H), 8.05-7.94 (m, 1H), 7.87 (d, J=7.0 Hz, 1H), 7.59 (d, J=7.9 Hz, 1H), 7.44-7.31 (m, 4H), 7.25 (d, J=7.9 Hz, 3H), 7.18 (d, J=7.8 Hz, 1H), 7.12 (d, J=7.7 Hz, 1H), 7.07 (t, J=7.6 Hz, 1H), 6.69-6.54 (m, 2H), 6.38 (d, J=10.1 Hz, 1H), 5.96 (d, J=9.0 Hz, 1H), 5.06-4.93 (m, 1H), 4.64-4.55 (m, 1H), 4.15 (d, J=12.8 Hz, 1H), 4.12-4.05 (m, 1H), 4.02 (d, J=5.1 Hz, 3H), 3.98-3.77 (m, 3H), 3.68 (d, J=5.0 Hz, 3H), 3.58 (d, J=14.6 Hz, 1H), 3.42 (d, J=13.3 Hz, 1H), 3.07 (s, 3H), 2.99 (d, J=13.4 Hz, 1H), 2.89-2.82 (m, 1H), 2.74 (d, J=3.3 Hz, 4H), 2.23-2.10 (m, 1H). HRMS (ESI) m/z: calcd for C48H50N9O5S+ [M+H]+, 864.3650; found, 864.3653.
Referring to the method of example 1, the target compound (SIAIS337089) was prepared by using Osimertinib derivative SIAIS337051 and intermediate LM (SIAIS255121). (yellow solid, 9.5 mg, yield 40%). 1H NMR (500 MHz, Methanol-d4) δ 8.46 (d, J=15.7 Hz, 1H), 8.28 (s, 1H), 8.08-8.00 (m, 2H), 7.96-7.85 (m, 1H), 7.60 (dd, J=15.1, 7.3 Hz, 1H), 7.56-7.47 (m, 1H), 7.37-7.25 (m, 4H), 7.07 (d, J=9.4 Hz, 1H), 6.67-6.57 (m, 1H), 6.48 (d, J=16.9 Hz, 1H), 5.84 (d, J=10.1 Hz, 1H), 5.03 (d, J=17.4 Hz, 1H), 4.25-4.11 (m, 1H), 4.05 (s, 3H), 3.95 (s, 3H), 3.76 (d, J=14.6 Hz, 1H), 3.65 (s, 1H), 3.51 (d, J=12.5 Hz, 1H), 3.26 (s, 4H), 2.98 (s, 3H), 2.86 (d, J=13.8 Hz, 1H), 2.80 (d, J=6.6 Hz, 3H), 2.72 (d, J=14.2 Hz, 1H), 2.37 (s, 2H), 2.27-2.18 (m, 1H), 1.91 (s, 3H). HRMS (ESI) m/z: calcd for C45H48N9O5+[M+H]+, 794.3773; found, 794.3775.
Referring to the method of example 1, the target compound (SIAIS337090) was prepared by using Osimertinib derivative SIAIS337051 and intermediate LM (SIAIS255127). (yellow solid, 8.7 mg, yield 33%). 1H NMR (500 MHz, Methanol-d4) δ 8.44 (d, J=18.1 Hz, 1H), 8.31 (s, 1H), 8.11-7.99 (m, 2H), 7.70-7.63 (m, 1H), 7.44-7.41 (m, 3H), 7.33-7.27 (m, 3H), 7.07 (d, J=5.6 Hz, 1H), 6.64 (t, J=13.5 Hz, 1H), 6.47 (d, J=16.8 Hz, 1H), 5.84 (d, J=10.1 Hz, 1H), 5.10 (s, 1H), 4.26 (d, J=10.9 Hz, 2H), 4.01 (s, 3H), 3.98 (s, 1H), 3.86 (s, 3H), 3.73 (d, J=13.6 Hz, 1H), 3.49-3.44 (m, 2H), 3.22-3.09 (m, 3H), 2.91 (s, 3H), 2.81 (s, 3H), 2.77-2.65 (m, 1H), 2.36-2.31 (m, 3H), 2.04 (s, 1H), 1.61 (s, 2H), 1.41 (s, 2H), 1.28-1.22 (m, 6H). HRMS (ESI) m/z: calcd for C49H56N9O5+ [M+H]+, 850.4399; found, 850.4394.
Referring to the method of example 1, the target compound (SIAIS262064) was prepared by using Dacomitinib derivative B and intermediate LM (SIAIS213135). (yellow solid, 8.9 mg, yield 35%). HRMS (ESI) m/z: calcd for C46H54ClFN9O5S+ [M+H]+, 898.3636; found, 898.3631.
Referring to the method of example 1, the target compound (SIAIS262071) was prepared by using Dacomitinib derivative B and intermediate LM (SIAIS264009). (yellow solid, 9.5 mg, yield 41%). HRMS (ESI) m/z: calcd for C46H53ClFN10O6+ [M+H]+, 895.3817; found, 895.3815.
Referring to the method of example 1, the target compound (SIAIS262110) was prepared by using Dacomitinib derivative C and intermediate LM (SIAIS171090). (yellow solid, 9.2 mg, yield 48%). HRMS (ESI) m/z: calcd for C39H39ClFN8O6+ [M+H]+, 801.2380; found, 801.2389.
Referring to the method of example 1, the target compound (SIAIS262112) was prepared by using Dacomitinib derivative C and intermediate LM (SIAIS171089). (yellow solid, 9.5 mg, yield 49%). HRMS (ESI) m/z: calcd for C41H43ClFN8O6+ [M+H]+, 829.2693; found, 829.2697.
Referring to the method of example 1, the target compound (SIAIS262113) was prepared by using Dacomitinib derivative C and intermediate LM (SIAIS171079). (yellow solid, 9.3 mg, yield 46%). HRMS (ESI) m/z: calcd for C42H45ClFN8O6+ [M+H]+, 843.2850; found, 843.2844.
Referring to the method of example 1, the target compound (SIAIS262114) was prepared by using Dacomitinib derivative C and intermediate LM (SIAIS171091). (yellow solid, 9.6 mg, yield 45%). HRMS (ESI) m/z: calcd for C43H47ClFN8O6+ [M+H]+, 857.3006; found, 857.3009.
Referring to the method of example 1, the target compound (SIAIS262115) was prepared by using Dacomitinib derivative C and intermediate LM (SIAIS171092). (yellow solid, 9.8 mg, yield 43%). HRMS (ESI) m/z: calcd for C44H49ClFN8O6+ [M+H]+, 871.3163; found, 871.3167.
Referring to the method of example 1, the target compound (SIAIS262116) was prepared by using Dacomitinib derivative C and intermediate LM (SIAIS255121). (yellow solid, 8.4 mg, yield 45%). HRMS (ESI) m/z: calcd for C42H43ClFN8O5+ [M+H]+, 793.3023; found, 793.3028.
Referring to the method of example 1, the target compound (SIAIS262117) was prepared by using Dacomitinib derivative C and intermediate LM (SIAIS255119). (yellow solid, 8.4 mg, yield 45%). HRMS (ESI) m/z: calcd for C43H45ClFN8O5+ [M+H]+, 807.3180; found, 807.3183.
Referring to the method of example 1, the target compound (SIAIS262118) was prepared by using Dacomitinib derivative C and intermediate LM (SIAIS1220099). (yellow solid, 9.6 mg, yield 35%). HRMS (ESI) m/z: calcd for C48H57ClFN8O6+ [M+H]+, 927.3789; found, 927.3784.
Referring to the method of example 1, the target compound (SIAIS337021) was prepared by using Dacomitinib derivative B and intermediate LM (SIAIS1220141). (yellow solid, 9.4 mg, yield 32%). HRMS (ESI) m/z: calcd for C49H52ClFN9O5S+ [M+H]+, 932.3479; found, 932.3474.
Referring to the method of example 1, the target compound (SIAIS337024) was prepared by using Dacomitinib derivative B and intermediate LM (SIAIS1221131). (yellow solid, 9.1 mg, yield 33%). HRMS (ESI) m/z: calcd for C49H53ClFN10O5+ [M+H]+, 915.3867; found, 915.3864.
Referring to the method of example 1, the target compound (SIAIS337025) was prepared by using Dacomitinib derivative B and intermediate LM (SIAIS1222121). (yellow solid, 9.7 mg, yield 45%). HRMS (ESI) m/z: calcd for C43H46ClFN9O7+ [M+H]+, 854.3187; found, 854.3181.
Referring to the method of example 1, the target compound (SIAIS337026) was prepared by using Dacomitinib derivative B and intermediate LM (SIAIS1222125). (yellow solid, 9.3 mg, yield 42%). HRMS (ESI) m/z: calcd for C46H52ClFN9O7+ [M+H]+, 896.3657; found, 896.3651.
Referring to the method of example 1, the target compound (SIAIS337027) was prepared by using Dacomitinib derivative B and intermediate LM (SIAIS1222149). (yellow solid, 8.8 mg, yield 39%). HRMS (ESI) m/z: calcd for C47H54ClFN9O7+ [M+H]+, 910.3813; found, 910.3811.
Referring to the method of example 1, the target compound (SIAIS337028) was prepared by using Dacomitinib derivative B and intermediate LM (SIAIS1222151). (yellow solid, 8.6 mg, yield 34%). HRMS (ESI) m/z: calcd for C48H56ClFN9O7+ [M+H]+, 924.3970; found, 924.3974.
Referring to the method of example 1, the target compound (SIAIS337029) was prepared by using Dacomitinib derivative B and intermediate LM (SIAIS1222127). (yellow solid, 8.2 mg, yield 31%). HRMS (ESI) m/z: calcd for C47H56ClFN9O6+[M+H]+, 896.4021; found, 896.4029.
Referring to the method of example 1, the target compound (SIAIS337035) was prepared by using Dacomitinib derivative B and intermediate LM (SIAIS1204061). (yellow solid, 8.5 mg, yield 42%). HRMS (ESI) m/z: calcd for C47H55ClFN10O6+[M+H]+, 909.3973; found, 909.3978.
Referring to the method of example 1, the target compound (SIAIS337036) was prepared by using Dacomitinib derivative B and intermediate LM (SIAIS1204063). (yellow solid, 8.7 mg, yield 43%). HRMS (ESI) m/z: calcd for C48H57ClFN10O6+[M+H]+, 923.4130; found, 923.4128.
Referring to the method of example 1, the target compound (SIAIS337037) was prepared by using Dacomitinib derivative B and intermediate LM (SIAIS299135). (yellow solid, 8.9 mg, yield 41%). HRMS (ESI) m/z: calcd for C51H62ClFN9O6S+ [M+H]+, 982.4211; found, 982.4218.
Referring to the method of example 1, the target compound (SIAIS337038) was prepared by using Dacomitinib derivative B and intermediate LM (SIAIS1220099). (yellow solid, 9.3 mg, yield 46%). HRMS (ESI) m/z: calcd for Cs2H64ClFN9O6S+ [M+H]+, 996.4367; found, 996.4362.
Referring to the method of example 1, the target compound (SIAIS337039) was prepared by using Dacomitinib derivative B and intermediate LM (SIAIS1220013). (yellow solid, 9.8 mg, yield 47%). HRMS (ESI) m/z: calcd for C51H64ClFN9O5S+ [M+H]+, 968.4418; found, 968.4418.
Referring to the method of example 1, the target compound (SIAIS337040) was prepared by using Dacomitinib derivative B and intermediate LM (SIAIS1220015). (yellow solid, 9.9 mg, yield 43%). HRMS (ESI) m/z: calcd for Cs2H66ClFN9O5S+ [M+H]+, 982.4575; found, 982.4575.
Referring to the method of example 1, the target compound (SIAIS262130) was prepared by using Caneritinib derivative A and intermediate LM (SIAIS151008). (white solid, 10.8 mg, yield 36%). HRMS (ESI) m/z: calcd for C55H75ClFN10O1S+ [M+H]+, 1173.5005; found, 1173.5001.
Referring to the method of example 1, the target compound (SIAIS249081) was prepared by using Dacomitinib derivative B and intermediate LM (SIAIS151025). (yellow solid, 8.6 mg, yield 54%). 1H NMR (500 MHz, Methanol-d4) δ 9.09 (s, 1H), 8.61 (s, 1H), 7.97 (dd, J=6.7, 2.6 Hz, 1H), 7.66 (ddd, J=8.9, 4.1, 2.5 Hz, 1H), 7.57 (dd, J=8.5, 7.1 Hz, 1H), 7.33-7.27 (m, 2H), 7.13 (d, J=7.1 Hz, 1H), 7.00 (dd, J=14.9, 7.5 Hz, 1H), 6.89 (d, J=8.4 Hz, 1H), 6.81 (d, J=15.1 Hz, 1H), 5.08 (dd, J=12.6, 5.5 Hz, 1H), 4.13 (s, 3H), 4.03 (s, 2H), 3.99 (d, J=7.2 Hz, 1H), 3.61-3.56 (m, 1H), 3.23-3.16 (m, 1H), 2.86 (ddd, J=17.8, 14.3, 5.2 Hz, 1H), 2.79-2.67 (m, 2H), 2.23-2.03 (m, 4H), 1.88-1.78 (m, 2H), 1.65-1.58 (m, 1H), 1.39-1.35 (m, 2H). HRMS (ESI) m/z: calcd for C39H38ClFN9O7+ [M+H]+, 798.2561; found, 798.2563.
Referring to the method of example 1, the target compound (SIAIS249082) was prepared by using Dacomitinib derivative B and intermediate LM (SIAIS151026). (yellow solid, 9.6 mg, yield 56%). 1H NMR (500 MHz, Methanol-d4) δ 9.24 (s, 1H), 8.62 (s, 1H), 7.99 (dd, J=6.7, 2.6 Hz, 1H), 7.74-7.67 (m, 1H), 7.59 (dd, J=8.6, 7.1 Hz, 1H), 7.36-7.26 (m, 2H), 7.15 (d, J=8.6 Hz, 1H), 7.06 (dd, J=14.6, 7.5 Hz, 2H), 6.80 (d, J=15.2 Hz, 1H), 5.10 (dd, J=12.3, 5.3 Hz, 1H), 4.14 (s, 3H), 3.98 (d, J=7.0 Hz, 2H), 3.90 (s, 1H), 3.75-3.66 (m, 1H), 3.09 (s, 1H), 2.94-2.82 (m, 1H), 2.80-2.69 (m, 2H), 2.58-2.51 (m, 2H), 2.22-2.03 (m, 4H), 1.65-1.58 (m, 2H), 1.38-1.28 (m, 3H). HRMS (ESI) m/z: calcd for C40H40ClFN9O7+ [M+H]+, 812.2718; found, 812.2713.
Referring to the method of example 1, the target compound (SIAIS249083) was prepared by using Dacomitinib derivative B and intermediate LM (SIAIS151019). (yellow solid, 8.7 mg, yield 51%). 1H NMR (500 MHz, Methanol-d4) δ 9.24 (s, 1H), 8.66 (s, 1H), 7.97 (dd, J=6.7, 2.7 Hz, 1H), 7.68 (ddd, J=8.9, 4.2, 2.7 Hz, 1H), 7.56 (dd, J=8.6, 7.1 Hz, 1H), 7.37-7.28 (m, 2H), 7.09 (d, J=8.5 Hz, 1H), 7.04 (dd, J=7.3, 4.0 Hz, 1H), 6.82 (d, J=15.2 Hz, 1H), 5.07 (dd, J=12.5, 5.4 Hz, 1H), 4.15 (s, 3H), 4.00 (d, J=7.2 Hz, 2H), 3.81-3.75 (m, 1H), 3.59-3.48 (m, 1H), 3.47-3.42 (m, 2H), 3.23-3.14 (m, 1H), 2.91-2.85 (m, 1H), 2.75 (dt, J=14.3, 3.1 Hz, 2H), 2.31 (d, J=8.4 Hz, 2H), 2.13-2.08 (m, 1H), 2.13-2.01 (m, 4H), 1.7-1.59 (m, 2H), 1.38-1.29 (m, 2H). HRMS (ESI) m/z: calcd for C41H42ClFN9O7+ [M+H]+, 826.2874; found, 826.2870.
Referring to the method of example 1, the target compound (SIAIS249084) was prepared by using Dacomitinib derivative B and intermediate LM (SIAIS151020). (yellow solid, 11.1 mg, yield 65%). 1H NMR (500 MHz, Methanol-d4) δ 9.17 (s, 1H), 8.64 (s, 1H), 7.98 (s, 1H), 7.68 (s, 1H), 7.55 (s, 1H), 7.32 (dd, J=19.0, 6.5 Hz, 2H), 7.1-7.00 (m, 3H), 6.82 (d, J=15.1 Hz, 1H), 5.07 (dd, J=12.5, 5.4 Hz, 1H), 4.15 (s, 3H), 4.00 (s, 1H), 3.62-3.5 (m, 1H), 3.21-3.15 (m, 1H), 2.84 (d, J=17.2 Hz, 1H), 2.75-2.71 (m, 2H), 2.28-2.18 (m, 2H), 2.09-1.99 (m, 4H), 1.72-1.68 (m, 4H), 1.63-1.58 (m, 1H) 1.48-1.38 (m, 2H), 1.35-1.28 (m, 4H). HRMS (ESI) m/z: calcd for C43H46ClFN9O7+ [M+H]+, 854.3187; found, 854.3189.
Referring to the method of example 1, the target compound (SIAIS249085) was prepared by using Dacomitinib derivative B and intermediate LM (SIAIS151086). (yellow solid, 10.6 mg, yield 59%). 1H NMR (500 MHz, Methanol-d4) δ 9.15 (s, 1H), 8.65 (s, 1H), 7.96 (dd, J=6.6, 2.6 Hz, 1H), 7.66 (ddd, J=8.9, 4.2, 2.6 Hz, 1H), 7.54 (dd, J=8.6, 7.1 Hz, 1H), 7.36-7.29 (m, 2H), 7.03 (dd, J=7.9, 2.6 Hz, 2H), 7.00 (t, J=7.4 Hz, 1H), 6.83 (d, J=15.2 Hz, 1H), 5.05 (dd, J=12.4, 5.5 Hz, 1H), 4.15 (s, 3H), 4.01 (d, J=7.2 Hz, 2H), 4.00-3.96 (m, 1H), 3.61-3.58 (m, 1H), 3.19-3.15 (m, 1H), 2.91-2.80 (m, 1H), 2.80-2.65 (m, 2H), 2.20 (q, J=7.6 Hz, 2H), 2.15-2.07 (m, 4H), 1.69-1.60 (m, 5H), 1.48-1.31 (m, 8H). HRMS (ESI) m/z: calcd for C44H48ClFN9O7+ [M+H]+, 868.3344; found, 868.3341.
Referring to the method of example 1, the target compound (SIAIS249086) was prepared by using Dacomitinib derivative B and intermediate LM (SIAIS151004). (yellow solid, 10.2 mg, yield 54%)1H NMR (500 MHz, Methanol-d4) δ 9.17 (s, 1H), 8.68 (s, 1H), 7.95 (dd, J=6.7, 2.6 Hz, 1H), 7.69-7.64 (m, 1H), 7.57-7.50 (m, 1H), 7.33 (dd, J=19.2, 10.3 Hz, 2H), 7.09 (d, J=8.5 Hz, 1H), 7.09-6.98 (m, 2H), 6.81 (d, J=15.2 Hz, 1H), 5.07 (dd, J=12.4, 5.5 Hz, 1H), 4.15 (s, 3H), 4.00 (d, J=7.2 Hz, 2H), 3.98-3.91 (m, 1H), 3.73 (dt, J=8.2, 5.7 Hz, 5H), 3.66-3.60 (m, 4H), 3.49 (t, J=5.3 Hz, 2H), 3.22-3.07 (m, 1H), 2.91-2.77 (m, 1H), 2.78-2.65 (m, 2H), 2.48-2.36 (m, 2H), 2.2-2.15 (m, 3H), 1.83-1.80 (m, 2H), 1.65-1.62 (m, 1H), 1.37-1.31 (m, 2H). HRMS (ESI) m/z: calcd for C44H48ClFN9O9+[M+H]+, 900.3242; found, 900.3241.
Referring to the method of example 1, the target compound (SIAIS249099) was prepared by using Caneritinib derivative A and intermediate LM (SIAIS164118). (yellow solid, 10.1 mg, yield 56%). HRMS (ESI) m/z: calcd for C45H49ClFN10O9+ [M+H]+, 927.3351; found, 927.3353.
Referring to the method of example 1, the target compound (SIAIS249100) was prepared by using Caneritinib derivative A and intermediate LM (SIAIS164119). (yellow solid, 9.6 mg, yield 59%). 1H NMR (500 MHz, Methanol-d4) δ 9.14 (s, 1H), 8.74 (s, 1H), 7.94 (dd, J=6.6, 2.6 Hz, 1H), 7.69-7.63 (mz, 1H), 7.54 (dd, J=8.6, 7.1 Hz, 1H), 7.42-7.33 (m, 2H), 7.12 (d, J=8.6 Hz, 1H), 7.02 (d, J=7.1 Hz, 1H), 6.78 (dd, J=16.9, 10.3 Hz, 1H), 6.51 (dd, J=16.9, 1.6 Hz, 1H), 5.90 (dd, J=10.2, 1.6 Hz, 1H), 5.05 (dd, J=12.7, 5.5 Hz, 2H), 4.48 (t, J=5.9 Hz, 2H), 3.71-3.61 (m, 3H), 3.52-3.40 (m, 6H), 2.93-2.82 (m, 1H), 2.77-2.66 (m, 4H), 2.70-2.63 (m, 6H), 2.13-2.05 (m, 1H), 2.05-2.01 (m, 1H), 1.39-1.36 (m, 2H). HRMS (ESI) m/z: calcd for C43H45ClFN10O8+ [M+H]+, 883.3089; found, 883.3081.
Referring to the method of example 1, the target compound (SIAIS249101) was prepared by using Caneritinib derivative A and intermediate LM (SIAIS1213011). (white solid, 8.2 mg, yield 37%). HRMS (ESI) m/z: calcd for C56H71ClFN12O7S+ [M+H]+, 1109.4956; found, 1109.4951.
Referring to the method of example 1, the target compound (SIAIS249102) was prepared by using Caneritinib derivative A and intermediate LM (SIAIS1213061). (white solid, 12 mg, yield 55%). 1H NMR (500 MHz, Methanol-d4) δ 9.16 (s, 1H), 9.12 (s, 1H), 8.75 (s, 1H), 7.93 (dd, J=6.6, 2.7 Hz, 1H), 7.7-7.64 (m, 1H), 7.47 (d, J=8.3 Hz, 1H), 7.42 (d, J=8.2 Hz, 2H), 7.37 (d, J=3.2 Hz, 2H), 7.15 (s, 4H), 6.84 (dd, J=16.9, 10.3 Hz, 1H), 6.50 (dd, J=16.9, 1.6 Hz, 1H), 5.89 (dd, J=10.3, 1.6 Hz, 1H), 4.61-4.53 (m, 3H), 4.51-4.46 (m, 2H), 4.37 (d, J=15.5 Hz, 1H), 4.09 (s, 1H), 3.89 (d, J=11.0 Hz, 1H), 3.78 (dd, J=10.9, 4.0 Hz, 1H), 3.45-3.40 (m, 3H), 3.30-3.19 (m, 1H), 2.94-2.82 (m, 5H), 2.79-2.72 (m, 3H), 2.64-2.53 (m, 3H), 2.49 (s, 3H), 2.28-2.17 (m, 1H), 2.12-1.99 (m, 1H), 1.29 (s, 2H), 0.96 (s, 9H). HRMS (ESI) m/z: calcd for C55H67ClFN10O7S+ [M+H]+, 1101.4582; found, 1101.4580.
Referring to the method of example 1, the target compound (SIAIS249103) was prepared by using Dacomitinib derivative C and intermediate LM (SIAIS164118). (yellow solid, 11.6 mg, yield 64%). 1H NMR (500 MHz, Methanol-d4) δ 9.27 (s, 1H), 8.75 (s, 1H), 7.92 (dd, J=6.7, 2.6 Hz, 1H), 7.64-7.68 (m, 1H), 7.60-7.55 (m, 1H), 7.37 (t, J=8.9 Hz, 1H), 7.32 (s, 1H), 7.11-7.02 (m, 3H), 6.87 (d, J=15.1 Hz, 1H), 5.09 (dd, J=12.1, 5.4 Hz, 1H), 4.18 (s, 3H), 4.08 (s, 2H), 3.79 (d, J=12.6 Hz, 2H), 3.72 (dt, J=9.0, 5.1 Hz, 3H), 3.67-3.63 (m, 2H), 3.58 (t, J=5.2 Hz, 2H), 3.53 (dt, J=6.0, 2.6 Hz, 2H), 3.49 (t, J=5.1 Hz, 2H), 3.48-3.38 (m, 3H), 3.26-3.18 (m, 1H), 2.89-2.82 (m, 1H), 2.79-2.70 (m, 3H), 2.70-2.45 (m, 6H), 2.28-2.17 (m, 2H), 2.15-2.10 (m, 2H), 1.38-1.32 (m, 2H). HRMS (ESI) m/z: calcd for C49H56ClFN11O9+ [M+H]+, 996.3930; found, 996.3931.
Referring to the method of example 1, the target compound (SIAIS249104) was prepared by using Dacomitinib derivative C and intermediate LM (SIAIS164119). (yellow solid, 11.8 mg, yield 70%). 1H NMR (500 MHz, Methanol-d4) δ 9.26 (s, 1H), 8.74 (s, 1H), 7.93 (dd, J=6.6, 2.6 Hz, 1H), 7.68-7.63 (m, 1H), 7.56 (dd, J=8.6, 7.1 Hz, 1H), 7.37 (t, J=8.9 Hz, 1H), 7.32 (s, 1H), 7.13 (d, J=8.6 Hz, 1H), 7.08-7.02 (m, 2H), 6.86 (d, J=15.2 Hz, 1H), 5.07 (dd, J=12.7, 5.5 Hz, 1H), 4.18 (s, 3H), 4.05 (s, 2H), 3.75 (s, 2H), 3.50-3.41 (m, 8H), 3.22-3.12 (m, 3H), 2.92-2.86 (m, 1H), 2.78-2.58 (m, 5H), 2.55-2.41 (m, 4H), 2.25-2.15 (m, 2H), 2.14-2.09 (m, 2H), 1.35-1.30 (m, 2H). HRMS (ESI) m/z: calcd for C47H52ClFN11O8+ [M+H]+, 952.3667; found, 952.3665.
Referring to the method of example 1, the target compound (SIAIS249105) was prepared by using Dacomitinib derivative C and intermediate LM (SIAIS1213011). (white solid, 12.1 mg, yield 57%). 1H NMR (500 MHz, Methanol-d4) δ 9.53 (s, 1H), 9.26 (s, 1H), 8.75 (s, 1H), 7.92 (dd, J=6.6, 2.6 Hz, 1H), 7.70-7.65 (m, 1H), 7.57-7.51 (m, 2H), 7.50-7.44 (m, 2H), 7.40-7.32 (m, 2H), 7.07 (dt, J=14.8, 7.2 Hz, 1H), 6.89 (d, J=15.2 Hz, 1H), 4.67-4.46 (m, 5H), 4.39 (dd, J=15.8, 2.0 Hz, 1H), 4.18 (s, 3H), 4.10 (s, 2H), 3.97 (d, J=11.0 Hz, 2H), 3.88-3.45 (m, 24H), 3.28-3.20 (m, 2H), 2.91-2.87 (m, 2H), 2.55 (s, 3H), 2.33-2.16 (m, 3H), 2.12-1.98 (m, 1H), 1.32-1.28 (m, 2H), 1.06 (s, 9H). HRMS (ESI) m/z: calcd for C60H78ClFN13O7S+ [M+H]+, 1178.5535; found, 1178.5531.
Referring to the method of example 1, the target compound (SIAIS249106) was prepared by using Dacomitinib derivative C and intermediate LM (SIAIS1213061). (white solid, 13 mg, yield 56%). 1H NMR (500 MHz, Methanol-d4) δ 9.26 (s, 1H), 9.14 (s, 1H), 8.75 (s, 1H), 7.92 (dt, J=6.6, 2.8 Hz, 1H), 7.68-7.62 (m, 1H), 7.49-7.45 (m, 2H), 7.44-7.41 (m, 1H), 7.39-7.31 (m, 2H), 7.15 (s, 3H), 7.07 (dt, J=14.7, 7.1 Hz, 1H), 6.88 (d, J=15.2 Hz, 1H), 4.65-4.45 (m, 4H), 4.38 (d, J=15.6 Hz, 1H), 4.18 (s, 3H), 4.09 (s, 2H), 3.89 (d, J=11.0 Hz, 1H), 3.79 (dd, J=10.9, 4.0 Hz, 3H), 3.62 (d, J=12.1 Hz, 2H), 3.29-3.16 (m, 4H), 2.92-2.81 (m, 5H), 2.73 (d, J=7.7 Hz, 2H), 2.63-2.53 (m, 2H), 2.49 (s, 3H), 2.48-2.45 (m, 1H), 2.29-2.16 (m, 3H), 2.13-2.00 (m, 1H), 1.37-1.27 (m, 3H), 0.97 (s, 9H), 0.95-0.83 (m, 2H). HRMS (ESI) m/z: calcd for C62H74ClFN11O7S+ [M+H]+, 1170.5160; found, 1170.5161.
Halt protease and phosphatase inhibitors (Thermo Fisher)
Cell TITER BLUE detection kit (Promega)
Cell TITER GLO detection kit (Promega)
Cell counting kit-8 (CCK-8; WST) (Dojindo)
Fetal bovine serum (GIBICO)
Most of the antibodies used for the following assays were purchased from Cell Signaling Technology Company, such as EGFR (#4267S) etc; and α-tubulin and GAPDH were purchased from Abcam Company.
The cell lines with abnormal EGFR expression used are as follows:
HCC827 cells (EGFR Exon19 del, non-small cell lung cancer cells);
PC9 cells (EGFR Exon 19 del, non-small cell lung cancer cells);
PC9BracA1 (EGFR Exon 19 del+T790M, non-small cell lung cancer cells)
H1975 (EGFR Exon T790M+L858R mutation, non-small cell lung cancer cell line)
BT474 cells (HER2 positive breast cancer cells)
The tumor cell lines used were routinely cultured in an incubator with 5% CO2 at 37° C.
HCC827 and H1975 cells were purchased from ATCC. Sources of PC9 and PC9BracA1 cells can be found in articles (Song, 2015) and (Chmielecki, 2011). The medium for non-small cell lung cancer cell lines is RPMI1640 supplemented with 10% FCS and Penicillin-Streptomycin. BT474 Breast cancer cells were cultured in RPMI1640 supplemented with 10% FCS, Insulin and Penicillin-Streptomycin. The cells used were identified as correct cells by STR cells, and were negative for mycoplasma through routine inspections.
Construction of EGFR triple mutant cell line PC9 DCT (Del19+T790M+C797S): The cDNA expressing human EGFR was cloned into pLVX vector, and exon 19 deletion mutation (746-750) and T790M mutation were introduced by point mutation and C797S mutation. Transferred into PC9 cells by lentiviral packaging. Cells stably expressing EGFR triple mutations were obtained by fluorescence screening.
Generation of EGFR triple mutant cell line PC9 DCT (Del19+T790M+C797S):
The cDNA sequence expressing human EGFR was cloned into a pLVX vector (GFP+), and then introduce the EGFR exon 19 deletion (746-750), exon 20 T790M and C797S mutations by were generated by site-directed mutagenesis. The PC9 cells were infected with lentivirus containing the indicated EGFR mutations, and the cells stably expressing EGFR triple mutation were obtained by fluorescence screening.
Cancer cells were planted in 12-well-plate at the density of 0.15 million cells per well with 1 mL RPMI1640 completed culture medium. Compounds with different concentrations (DMSO as a solution system, and 1 μL of compounds of different concentrations was added to 1 mL of the cell culture medium) were added to the cells planted one day before. 16 hours after treated, the medium was discarded and the cells were washed with precooled PBS. The cells were placed on ice and treated with RIPA protein lysis buffer containing halt protease and phosphatase inhibitor. After centrifugation at 10000 rpm at 4° C. for 10 minutes, the supernatant was collected. The same amount of proteins were added into 4×SDS loading buffer and heated at 95° C. for 5 minutes for denaturation, and the samples were stored at −20° C. after denaturation, or directly conducted protein electrophoresis. The 4-20% gradient preformed Electrophoretic gels were purchased from Kingsy. The electrophoresis tank and related components were purchased from Bio-rad company, and the electrophoresis condition was isobaric 120V for 2 hours. PVDF membrane was used for the transmembrane, and the whole transmembrane process was carried out on ice with a constant current of 0.4 amperes for an hour. The membrane was blocked with 5% skimmed milk dilution with TBST buffer. Refer to the corresponding antibody instructions for the specific steps of immunoblotting.
All of the half inhibitory concentrations (IC50) of these compounds of the present invention were determined with WST reagent from Fuyuan Biotec Company. The specific experiment procedures are as follows: Cancer cells were planted in 96-well-plate at the density of 3 000 cells per well with 100 μL RPMI1640 completed culture medium. In the next day, compounds with indicated concentrations were added to the cells, DMSO was used as the negative control and the EGFR TKIs were used to treat cells as the positive groups. After 72 hours drug treatment, cell viability was measured by using the WST reagent following the instructions. The growth inhibition curve of the compounds on cells were plotted by Prism GraphPad, and the compounds IC50 calculated from it. The specific results are shown in the tables.
The concentration-dependent assays were taken to determine the anti-tumor activity of these indicated compounds in HCC827 and PC9 cell lines, both of which have the EGFR Exon 19 deletion mutation, sensitive to EGFR inhibitors. The cells were treated with the gradient concentration of these compounds (the maximum concentration: 10 μM; 3-fold dilution ratio, 10 concentration gradients) for 72 hours and then incubated with the WST (CCK8) reagent to determine the cell viability. The results are shown as follows:
As shown in Table 1, all of the dacomitinib-based compounds of the present invention can inhibit the cell growth of HCC827 or PC9 cells well. The IC50 values of some compounds of the present invention are close to which of dacomitinib on HCC827 cells (0.5 nM), for instance, the IC50 of SIAIS262033 is about 1.0 nM. The cell killing effect of dacomitinib on PC9 cells (IC50: 0.4 nM) is higher than that of the dacomitinib derivatives we synthesized, while some of the compounds of the present invention have the similar efficacy as dacomitinib, such as SIAIS249059 with the IC50 1.2 nM on PC9 cells.
Moreover, the cell viability assays were taken on H1975 cells as well, and the results indicated that these compounds of the present invention also showed cell proliferation inhibition effect in H1975 and the detailed results are shown in Table 2.
1.2 Studies on the decrease of EGFR protein level caused by the dacomitinib-based compounds of the present invention
The degradation efficacy of the dacomitinib-based compounds on EGFR was determined in HCC827 and H1975 cell lines, which harboring EGFR Exon 19del and EGFR L858R+T790M mutations respectively.
Dacomitinib with gradient concentrations (1, 10, 50, 100, 500 nM) were used to treat HCC827 cells for 16 hours, then the western blot assay was taken to estimate the EGFR levels of the cell lysates in each group. As shown in
As shown in
In
The cell viability assays were taken to determine the anti-tumor activity of these indicated compounds of the present invention in HCC827 and PC9 cell lines, both of which have the EGFR Exon 19 deletion mutation, sensitive to EGFR inhibitors. The cells were treated with the gradient concentration of these compounds (the maximum concentration: 10 μM; 3-fold dilution ratio, 10 concentration gradients) for 72 hours and then incubated with the WST (CCK8) reagent to determine the cell viability. The results are shown as Table 3.
All poziotinib-based compounds of the present invention can inhibit the cell proliferation of HCC827 and PC9 cells well (Table 3). The IC50 values of the compounds we synthesized are close to which of poziotinib on HCC827 cells (0.5 nM).
The cell viability assays were taken to determine the anti-tumor activity of these indicated compounds in HCC827 cells with the EGFR Exon 19 deletion mutation, which is sensitive to EGFR inhibitors. The cells were treated with the gradient concentration of these compounds (the maximum concentration: 10 μM; 3-fold dilution ratio, 10 concentration gradients) for 72 hours and then incubated with the WST reagent to determine the cell viability. The experiments were repeated more than 3 times and the results are shown as Table 4.
The cell proliferation inhibitory efficacy of gefitinib derivatives A or B-based PROTAD compounds of the present invention on HCC827 cells is weaker than that of gefitinib with the IC50 4.8 nM, while the gefitinib derivative C-based PROTAD compounds could inhibit the HCC827 cell growth well, for instance the SIAIS293052 has the similar cell killing capacity as gefitinib. All of the sapitinib (AZD8931)—based compounds could inhibit cell proliferation well, such as SIAIS293067 which is better than sapitinib on cell killing.
The cell viability assays were taken to determine the anti-tumor activity of these indicated compounds in HCC827 cells with the EGFR Exon 19 deletion mutation, which is sensitive to EGFR inhibitors. The cells were treated with the gradient concentration of these compounds (the maximum concentration: 10 μM; 3-fold dilution ratio, 10 concentration gradients) for 72 hours and then incubated with the WST reagent to determine the cell viability. The results are shown as Table 5.
All series of afatinib derivative-based compounds of the present invention could inhibit the proliferation of HCC827 cells obviously (Table 5), and the inhibitory efficacies were similar as that of afatinib with the IC50 of 1.1 nM.
The cell viability assays were taken to determine the anti-tumor activity of these indicated compounds in HCC827 and PC9 cell lines, both of which have the EGFR Exon 19 deletion mutation, sensitive to EGFR inhibitors. The cells were treated with the gradient concentration of these compounds (the maximum concentration: 10 μM; 3-fold dilution ratio, 10 concentration gradients) for 72 hours and then incubated with the WST reagent to determine the cell viability. The results are shown as Table 6.
All of the canertinib-based compounds of the present invention can inhibit the cell proliferation of HCC827 and PC9 cells well (Table 6). The IC50 values of canertinib on HCC827 and PC9 cells are 0.7 nM and 0.9 nM, respectively, and the canertinib-based PROTAD compounds of the present invention showed the inhibitory effect equivalent to that of canertinib, for instance, the half-inhibition concentration of SIAIS262125 in HCC827 cells is 0.3 nM.
The cell viability assays were also taken to determine the cell proliferation inhibitory ability of these compounds in H1975 and PC9Brcal NSCLC cell lines, both of which have the acquired EGFR T790M drug resistance mutation against the 1st generation EGFR TKIs. The results reflected that these compounds of the present invention could inhibit the cell proliferation of H1975 and PC9Brcal cells well (Table 7), and most of the PROTAD compounds showed greater capacity in cell growth inhibition than canertinib. The IC50 values of SIAIS262125 and SIAIS262182 on H1975 cell are less than 25 nM, which means that the development of degradation agents to overcome T790M mutation is promising.
The degradation efficacy of the canertinib-based compounds on EGFR was determined in H1975 cells with the EGFR L858R+T790M mutation.
The H1975 cells were treated with the PROTADs with different concentrations (1, 10, 50, 100, 500 nM) for 16 hours, and then the western blot assay was taken to estimate the EGFR levels of the cell lysates in each group. As shown in
Moreover, the effect on EGFR phosphorylation level caused by the compounds of the present invention was also studied (
The cell viability assays were taken to determine the anti-tumor activity of these indicated compounds in HCC827, PC9, H1975 and PC9Bracl cell lines, which have the EGFR Exon 19 deletion mutation, sensitive to EGFR inhibitors. The cells were treated with the gradient concentration of these compounds (the maximum concentration: 10 μM; 3-fold dilution ratio, 10 concentration gradients) for 72 hours and then incubated with the WST reagent to determine the cell viability. The results are shown as Table 8.
7. Studies on the Anti-Tumor Capacity of the PROTAD Compounds of the Present Invention in Cells with EGFR Tertiary Drug Resistance Mutations.
The cell viability assays were taken to determine the anti-tumor activity of these indicated compounds in PC9 cells, which overexpressed Exon 19del+T790M+C797S triple mutant EGFR. The results showed that the poziotinib-based PROTADs could inhibit the PC9 drug resistant cells proliferation much better than the poziotinib derivative.
The degradation efficacy of some of the PROTAD compounds of the present invention on EGFR was determined in PC9 cells harboring EGFR triple mutations (19del+T790M+C797S). The results reflected that these compounds of the present invention could degrade the mutant EGFR effectively, suggesting that these PROTADs have great anti-tumor efficacy in drug resistant PC9 cells with EGFR triple mutations.
The cell viability assays were taken to determine the anti-tumor activity of these indicated compounds in BT474 cells, which can overexpress HER2 protein. The cells were treated with the gradient concentration of these compounds (the maximum concentration: 10 μM; 3-fold dilution ratio, 10 concentration gradients) for 72 hours and then incubated with the CCK8 reagent to determine the cell viability. The results are shown in Table 10. Some of these PROTADs displayed better antineoplastic activity than the relative EGFR TKIs, such as compound SIAIS262125 etc.
Meanwhile, the HER2 levels in BT474 cells treated with the compounds of the present invention were determined and the results indicated that these compounds of the present invention have the ability of HER2 degradation at high concentrations (
In summary, the PROTAD compounds of the present invention can effectively overcomes the various shortcomings in the prior small molecular targeted therapy on lung cancers and has high industrial utilization value.
The basic principles, main features and advantages of the present disclosure are shown and described above. Those skilled in the art should understand that the present disclosure is not limited by the foregoing embodiments, and they can make various changes, substitutions and alterations herein without departing from the spirit and scope of the present disclosure. These changes, substitutions and alterations fall within the scope of the present disclosure. The claimed scope of the present disclosure is defined by the appended claims and their equivalents.
| Number | Date | Country | Kind |
|---|---|---|---|
| 201910717328.8 | Aug 2019 | CN | national |
This application is the U.S. national phase of International Application No. PCT/CN2020/107177 filed Aug. 5, 2020 which designated the U.S. and claims priority to CN Patent Application No. 201910717328.8 filed Aug. 5, 2019, the entire contents of each of which are hereby incorporated by reference.
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/CN2020/107177 | 8/5/2020 | WO |
