Patent Application
20190322698
The present invention relates to the field of medicines, and particularly to a class of water-soluble epiallopregnanolone (3β,5α-tetrahydroprogesterone) derivatives, a pharmaceutical composition comprising the same, and use thereof in prevention or treatment of central nervous system diseases, in treatment of Alzheimers disease, in treatment of epilepsy or in treatment of depression.
Neurosteroids play an important physiological role in human body. Damages to the synthesis of neurosteroids in vivo will cause different neurological or mental diseases (1. Expert Opin Ther Targets. 2014, 18(6):679-90; 2. Neuroscience 2011, 191:55-77). There are a wide range of neurosteroids, among which epiallopregnanolone (3β,5α-tetrahydroprogesterone) is a GABA (γ-aminobutyric acid) receptor modulating neurosteroid antagonist (Brain Research 2003, 982:45-53), which is useful in prevention or treatment of central nervous system diseases (WO9945931A1 and CN102753181B), in treatment of premenstrual syndrome, in treatment of premenstrual dysphoric disorder, in treatment of menstrual-related migraine, in treatment of Alzheimers disease, in treatment of epilepsy, or in treatment of depression, etc.
As a steroid compound, epiallopregnanolone has lower solubility in water (CN102753181B). Therefore, it is quite necessary to develop epiallopregnanolone derivatives having improved water solubility and safety.
In one aspect, the present invention provides a water-soluble epiallopregnanolone derivative. The water-soluble epiallopregnanolone derivative has good physical/chemical stability and good water solubility, and furthermore can be dissociated rapidly in vivo to release the active drug (epiallopregnanolone), thus providing pharmacological effects quickly.
The water-soluble epiallopregnanolone derivative of the present invention is a compound of formula (I), or a pharmaceutically acceptable salt, a stereoisomer, a polymorph, or a solvate thereof:
wherein
X is H or F;
Y is H, F, or C1-6 alkyl optionally substituted with one or more F;
n is 0, 1, 2, 3, 4, 5 or 6;
W is W1 or W2;
W1 is NR1R2.A or
R1 and R2 are each independently H, C1-6 alkyl optionally substituted with phenyl, or C3-6 cycloalkyl;
m is 0, 1, 2 or 3;
A is absent or is a pharmaceutically acceptable acid;
W2 is —COOH, —OPO3(H)2, —PO3(H)2, —COO(M)1/t, —OPO3(M)2/t or —PO3(M)2/t;
M is a metal ion, an ammonium ion or a basic amino acid cation; and
t is the charge number of M.
In a further aspect, the present invention provides a pharmaceutical composition comprising a prophylactically or therapeutically effective amount of the compound of formula (I) of the present invention, or a pharmaceutically acceptable salt, a stereoisomer, a polymorph, or a solvate thereof, and one or more pharmaceutically acceptable carriers.
In a further aspect, the present invention provides use of the compound of formula (I) of the present invention or a pharmaceutically acceptable salt, a stereoisomer, a polymorph, or a solvate thereof or the pharmaceutical composition of the present invention in the manufacture of a medicament for preventing or treating central nervous system diseases, for treating Alzheimers disease, for treating epilepsy, or for treating depression.
In a further aspect, the present invention provides the compound of formula (I) of the present invention, or a pharmaceutically acceptable salt, a stereoisomer, a polymorph, or a solvate thereof, or the pharmaceutical composition of the present invention, for use in prevention or treatment of central nervous system diseases, in treatment of Alzheimer s disease, in treatment of epilepsy, or in treatment of depression.
In a further aspect, the present invention provides a method for preventing or treating central nervous system diseases, for treating Alzheimers disease, for treating epilepsy, or for treating depression, comprising administering a prophylactically or therapeutically effective amount of the compound of formula (I) of the present invention, or a pharmaceutically acceptable salt, a stereoisomer, a polymorph, or a solvate thereof, or the pharmaceutical composition of the present invention to a subject in need thereof.
Unless otherwise defined below, all technical and scientific terms and intents used herein should be identical to those commonly understood by those skilled in the art. Techniques used herein refer to those techniques commonly understood in the art, including apparent variations and replacement of equivalent techniques for those skilled in the art. Although we believe the following terms will be well understood by those skilled in the art, the definitions of which are provided herein to better illustrate the present invention.
As used herein, the terms “comprising”, “including”, “having”, “containing” or “relating to” and other variations in this context are inclusive or open-ended and do not exclude other elements, method, or steps not listed.
As used herein, the term “C1-6 alkyl” refers to a saturated, linear or branched hydrocarbon group having 1-6 (e.g., 1, 2, 3, 4, 5 or 6) carbon atoms, such as methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neo-pentyl, n-hexyl, isohexyl, etc., preferably methyl, ethyl, propyl, isopropyl, butyl or isobutyl, and more preferably methyl, ethyl or propyl.
As used herein, the term “C3-6 cycloalkyl” refers to a saturated monocyclic hydrocarbon group having 3-6 (e.g., 3, 4, 5 or 6) carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.
As used herein, the term “C6-14 aryl” refers to an aromatic group having 6 to 14 carbon atoms, such as phenyl or naphthyl.
The pharmaceutically acceptable salts of the compound of the present invention include the salts formed from the compound and pharmaceutically acceptable acids, and the salts formed from the compound and pharmaceutically acceptable bases.
As used herein, the term “pharmaceutically acceptable acid” refers to an acid that can be used in pharmaceuticals, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, carbonic acid, formic acid, acetic acid, acetoacetic acid, trifluoroacetic acid, propionic acid, pyruvic acid, butyric acid, caproic acid, heptanoic acid, undecanoic acid, lauric acid, stearic acid, palmitic acid, oxalic acid, methanesulfonic acid, trifluoromethanesulfonic acid, ethanedisulfonic acid, isethionic acid, 1,5-naphthalenedisulfonic acid, 2-naphthalenesulfonic acid, camphorsulfonic acid, sulfamic acid, lactic acid, benzenesulfonic acid, p-toluenesulfonic acid, malonic acid, succinic acid, glutaric acid, adipic acid, maleic acid, fumaric acid, lactic acid, tartaric acid, citric acid, malic acid, benzoic acid, salicylic acid, cinnamic acid, naphthoic acid, pamoic acid, nicotinic acid, orotic acid, methylsulfuric acid, dodecylsulfuric acid, glutamic acid, aspartic acid, gluconic acid, glucuronic acid, or any combination thereof.
As used herein, the term “pharmaceutically acceptable base” refers to a base that can be used in pharmaceuticals, such as inorganic bases (e.g., alkali metal hydroxide or alkaline earth metal hydroxide, etc.) or organic bases (e.g., amine (primary amine, secondary amine or tertiary amine), etc.). Examples of appropriate salts include, but not limited to, organic salts derived from amino acid, ammonia, primary amine, secondary amine, tertiary amine and cyclamine (e.g., diethylamine salt, piperidine salt, morpholine salt, piperazine salt, choline salt, meglumine salt, tromethamine salt, etc.) and inorganic salts derived from Na, Ca, K, Mg, Mn, Fe, Cu, Zn, Al and Li.
As used herein, the term “basic amino acid” refers to an amino acid in which OH− generated by hydrolysis is more than H+, e.g., arginine, lysine or histidine.
As used herein, the term “stereoisomer” represents isomers formed due to at least one asymmetric center. A compound having one or more (e.g., one, two, three or four) asymmetric centers may exist in the form of a racemic mixture, a single enantiomer, a diastereomeric mixture and a single diastereoisomer.
The compound of the present invention may exist in the form of crystal or polymorph, and may be a single polymorph or a mixture of more than one polymorph in any proportion.
The compound of the present invention may exist in the form of a solvate, especially a hydrate thereof, wherein the compound of the present invention comprises a polar solvent, e.g., water, ethanol, isopropanol, ethyl acetate or acetone, as a structural element of the crystal lattice. The polar solvent, especially water, may exist in a stoichiometric amount or a non-stoichiometric amount.
According to an embodiment of the present invention, provided is a water-soluble epiallopregnanolone derivative, which is a compound of formula (I), or a pharmaceutically acceptable salt, a stereoisomer, a polymorph, or a solvate thereof:
wherein
X is H or F;
Y is H, F, or C1-6 alkyl optionally substituted with one or more F;
n is 0, 1, 2, 3, 4, 5 or 6;
W is W1 or W2;
W1 is NR1R2.A or
R1 and R2 are each independently H, C1-6 alkyl optionally substituted with phenyl, or C3-6 cycloalkyl;
m is 0, 1, 2 or 3;
A is absent or is a pharmaceutically acceptable acid;
W2 is —COOH, —OPO3(H)2, —PO3(H)2, —COO(M)1/t, —OPO3(M)2/t or —PO3(M)2/t;
M is a metal ion, an ammonium ion or a basic amino acid cation; and
t is the charge number of M.
According to an embodiment of the present invention, Y is F, CF3, CH2F or CHF2.
According to an embodiment of the present invention, W is W1.
According to an embodiment of the present invention, R1 and R2 are each independently C1-6 alkyl optionally substituted with phenyl, e.g., methyl, ethyl, propyl, isopropyl, butyl, isobutyl or benzyl.
According to an embodiment of the present invention, R1 and R2 are each independently C3-6 cycloalkyl, e.g., cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.
According to an embodiment of the present invention, R1 and R2 are each independently H, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, benzyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.
According to an embodiment of the present invention, R1 and R2 are not H at the same time.
According to an embodiment of the present invention, W1 is selected from the group consisting of:
According to an embodiment of the present invention, A is hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, carbonic acid, formic acid, acetic acid, propionic acid, acetoacetic acid, trifluoroacetic acid, propionic acid, pyruvic acid, butyric acid, caproic acid, heptanoic acid, undecanoic acid, lauric acid, stearic acid, palmitic acid, oxalic acid, methanesulfonic acid, trifluoromethanesulfonic acid, ethanesulfonic acid, ethanedisulfonic acid, isethionic acid, 1,5-naphthalenedisulfonic acid, 2-naphthalenesulfonic acid, camphorsulfonic acid, sulfamic acid, lactic acid, benzenesulfonic acid, p-toluenesulfonic acid, malonic acid, succinic acid, glutaric acid, adipic acid, maleic acid, fumaric acid, tartaric acid, citric acid, malic acid, benzoic acid, salicylic acid, cinnamic acid, naphthoic acid, pamoic acid, nicotinic acid, orotic acid, methylsulfuric acid, dodecylsulfuric acid, glutamic acid, aspartic acid, gluconic acid, glucuronic acid, or any combination thereof.
According to an embodiment of the present invention, W is W2.
According to an embodiment of the present invention, M is an alkali metal ion (e.g., a lithium ion, a sodium ion or a potassium ion), an alkaline earth metal ion (e.g., a magnesium ion, a zinc ion or a calcium ion) or a trivalent metal ion (e.g., an aluminum ion).
According to an embodiment of the present invention, M is an ammonium ion represented by formula (NR3R4R5R6)+ or
wherein R3, R4, R5 and R6 are each independently H, alkyl optionally substituted with phenyl, cycloalkyl, or aryl; and p is 0, 1, 2 or 3.
In a preferred embodiment, R3, R4, R5 and R6 are each independently H, C1-6 alkyl optionally substituted with phenyl, C3-6 cycloalkyl, or C6-14 aryl. In a more preferred embodiment, R3, R4, R5 and R6 are each independently H, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, phenyl, benzyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl. In a most preferred embodiment, R3 and R4 are each independently H, methyl or ethyl.
In a preferred embodiment, M is selected from
According to an embodiment of the present invention, M is arginine+H+, lysine+H+ or histidine+H+.
In a preferred embodiment, W2 is selected from the group consisting of —COONa, —COOK, —P(═O)(OLi)2, —P(═O)(ONa)2, —P(═O)(OK)2, —P(═O)O2Mg, —OP(═O)(ONa)2 and —OP(═O)O2Ca.
In an embodiment, X is different from Y, and the carbon atom to which both X and Y are attached is in a single R configuration, in a single S configuration, or in a mixture of R and S configurations.
The present invention covers compounds obtained by any combination of the embodiments.
According to an embodiment of the present invention, variable R in the compound of the present invention is selected from the group consisting of:
According to a further embodiment of the present invention, provided is a pharmaceutical composition comprising a prophylactically or therapeutically effective amount of the compound of formula (I) of the present invention, or a pharmaceutically acceptable salt, a stereoisomer, a polymorph, or a solvate thereof, and one or more pharmaceutically acceptable carriers.
The term “pharmaceutically acceptable carrier” in the present invention refers to a diluent, an adjuvant, an excipient or a vehicle which is administrated together with a therapeutic agent, and within the scope of sound medical judgment is suitable for contact with humans and/or other animal tissues without undue toxicity, irritation, allergic reactions or with a reasonable benefit/risk ratio relative to its problems or complications.
The pharmaceutically acceptable carrier in the pharmaceutical composition of the present invention include, but not limited to, sterile liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like. When the pharmaceutical composition is administered intravenously, water is an exemplary carrier. Saline, glucose, and glycerol solution can also be used as liquid carriers, particularly for injectable solutions. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, maltose, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene glycol, water, ethanol, etc. The composition, if desired, may also contain minor amounts of wetting agents, emulsifying agents (e.g., Tween-80, etc.), or pH buffering agents.
The pharmaceutical composition of the present invention may be administrated via suitable routes. Preferably, the pharmaceutical composition of the present invention is administrated orally, intravenously, intraarterially, subcutaneously, intraperitoneally, intramuscularly or transdermally.
The composition of the present invention may be administered in an appropriate dosage form for these administration routes.
The dosage forms include, but not limited to, tablets, capsules, troches, dragees, powders, sprays, creams, ointments, suppositories, gels, pastes, lotions, ointments, aqueous suspensions, injectable solutions, elixirs and syrups.
The compound of the present invention can be used for treating a variety of diseases that can be treated by administering epiallopregnanolone.
A further embodiment of the present invention provides use of the compound of formula (I) of the present invention or a pharmaceutically acceptable salt, a stereoisomer, a polymorph, or a solvate thereof or the pharmaceutical composition of the present invention in the manufacture of a medicament for preventing or treating central nervous system diseases, for treating Alzheimers disease, for treating epilepsy, or for treating depression.
A further embodiment of the present invention provides the compound of formula (I) of the present invention or a pharmaceutically acceptable salt, a stereoisomer, a polymorph, or a solvate thereof, or the pharmaceutical composition of the present invention, for use in prevention or treatment of central nervous system diseases, in treatment of Alzheimers disease, in treatment of epilepsy, or in treatment of depression.
A further embodiment of the present invention provides a method for preventing or treating central nervous system diseases, for treating Alzheimers disease, for treating epilepsy, or for treating depression, comprising administering a prophylactically or therapeutically effective amount of the compound of formula (I) of the present invention, or a pharmaceutically acceptable salt, a stereoisomer, a polymorph, or a solvate thereof, or the pharmaceutical composition of the present invention to a subject in need thereof.
In a further embodiment of the present invention, the central nervous system diseases are selected from the group consisting of traumatic brain injury, essential tremor, epilepsy (including refractory status epilepticus and rare genetic epilepticus (e.g., Dravet syndrome and Rett syndrome)), depression, and Alzheimers disease.
The term “therapeutically effective amount” as used herein refers to an amount of the compound which will alleviate one or more symptoms of the diseases to be treated to a certain extent after administration.
Dosage regimens may be adjusted to provide the optimum desired response. For example, a single bolus may be administered, several divided doses may be administered over time, or the dose may be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation. It is to be noted that dosage values may vary with the type and severity of the condition to be alleviated, and may include single or multiple doses. It is to be further understood that for any particular subject, specific dosage regimens should be adjusted over time according to the individual need and the professional judgment of the person administering or supervising the administration of the composition.
The amount of the compound of the present invention administered will be dependent on the subject being treated, the severity of the disorder or condition, the rate of administration, the disposal of the compound and the discretion of the prescribing physician.
Unless otherwise indicated, the term “treating” or “treatment”, as used herein, means reversing, alleviating, inhibiting the progress of, or preventing the disorder or condition to which such term applies, or one or more symptoms of such disorder or condition.
As used herein, the term “subject” includes a human or a non-human animal. An exemplary human subject includes a human subject having a disease (such as one described herein) (referred to as a patient), or a normal subject. The term “non-human animal” as used herein includes all vertebrates, such as non-mammals (e.g. birds, amphibians, reptiles) and mammals, such as non-human primates, livestock and/or domesticated animals (such as sheep, dog, cat, cow, pig and the like).
The water-soluble epiallopregnanolone derivative of the present invention, which is obtained by structural modification of the hydroxy of epiallopregnanolone while retaining the pharmacological activity of epiallopregnanolone, has good physical/chemical stability. After injection of an aqueous solution comprising the epiallopregnanolone derivatives, the active drug is released in vivo, thus exerting pharmacological effects. Surprisingly, the water-soluble epiallopregnanolone derivative of the present invention has good water solubility, and thus can be formulated into appropriate liquid preparations. Furthermore, the water-soluble epiallopregnanolone derivative of the present invention can be easily dissociated in vivo to release the active drug. Therefore, the water-soluble epiallopregnanolone derivative of the present invention can improve the water solubility of epiallopregnanolone, thus reducing the side effects caused by adjuvants.
The present invention will be further described in detail with reference to the following examples for apparency of the purpose and technical solution of the present invention. It should be understood that these examples are provided merely for further illustration of the present invention, but should not be construed as limitation to the scope of the present invention. Any non-essential modifications and/or adjustments to the technical solutions of the present invention by a person skilled in the art based on the above disclosure of the present invention all fall within the protection scope of the present invention.
4-(Dimethylamino)-2-fluorobutyric acid hydrochloride (10 mmol) was placed in thionyl chloride (10 ml), followed by slowly heating to 40° C., and the mixture was allowed to react for 4 hours. Thionyl chloride was removed by evaporation under reduced pressure, and anhydrous dichloromethane (DCM) (15 ml) was added. After stirring, the solvent was removed by evaporation under reduced pressure. Anhydrous dichloromethane (20 ml) was added to the residues, and the solution thus obtained was used for the next step.
Epiallopregnanolone (4.5 mmol) and 4-dimethylaminopyridine (DMAP) (8.2 mmol) were dissolved in anhydrous dichloromethane (20 ml) at −78° C., and then a dichloromethane solution of intermediate 1 prepared in step 1) was slowly added dropwise. The reaction was monitored by HPLC. After the reaction was completed, the DCM layer was washed with an aqueous hydrochloric acid solution (pH=about 1.0), and the organic layer was dried over anhydrous sodium sulfate and filtered. The solvent was removed by rotary evaporation, and the residues were purified by preparative chromatography to obtain compound 1. Mass (ESI) [M-Cl]+=450.31.
At room temperature, epiallopregnanolone (10 mmol), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (80 mmol), (R)-4-(dimethylamino)-2-fluorobutyric acid hydrochloride (70 mmol) and DMAP (1 mmol) were added to anhydrous dichloromethane, and the mixture was heated to reflux under the protection of nitrogen. The reaction was monitored by HPLC. After the reaction was completed, the reaction solution was cooled to room temperature, and was successively washed with an aqueous hydrochloric acid solution (pH=3, 70 ml*3) and a saturated brine (70 ml*1). The organic layer was dried over anhydrous sodium sulfate and filtered. The solvent was removed by rotary evaporation, and the residues were recrystallized with isopropanol to obtain compound 2. Mass (ESI) [M-Cl]+=450.30.
Compound 3 was prepared according to the procedures similar to those in Example 1, except that 4-(dimethylamino)-2-fluorobutyric acid hydrochloride was replaced with (S)-4-(dimethylamino)-2-fluorobutyric acid hydrochloride (10 mmol) in step 1). Mass (ESI) [M-Cl]+=450.31.
Epiallopregnanolone (R)-5-(diethyl amino)-2-(trifluoromethyl)pentanoate hydrochloride was prepared according to the procedures similar to those in Example 1, except that 4-(dimethylamino)-2-fluorobutyric acid hydrochloride was replaced with (R)-5-(diethylamino)-2-(trifluoromethyl)pentanoic acid hydrochloride (10 mmol) in step 1). The hydrochloride salt was dissolved in dichloromethane, and the resulting organic phase was washed with an aqueous solution containing sodium methanesulfonate (adjusted to pH 3 with methanesulfonic acid). The organic layer was dried and filtered, and the solvent was removed by evaporation to obtain compound 6. Mass (ESI) [M-CH3SO3]+=514.32.
Epiallopregnanolone (4.5 mmol) and 4-dimethylaminopyridine (8.2 mmol) were dissolved in anhydrous dichloromethane (20 ml) at room temperature, and then dicyclohexyl carbodiimide (10 mmol) and 3-(dimethylamino)-2-fluoropropionic acid hydrochloride (8 mmol) were added. The mixture was allowed to react at room temperature, and the reaction was monitored by HPLC. After the reaction was completed, the DCM layer was washed with an aqueous hydrochloric acid solution (pH=about 1.0), and the organic layer was dried over anhydrous sodium sulfate and filtered. The solvent was removed by rotary evaporation, and the residues were purified by preparative chromatography to obtain compound 7. Mass (ESI) [M-Cl]+=436.30.
Epiallopregnanolone (4.5 mmol) and 4-dimethylaminopyridine (8.2 mmol) were dissolved in anhydrous dichloromethane (20 ml) at −40° C., and a dichloromethane solution of benzyl (R)-4-chloro-3-fluoro-4-oxobutyrate (prepared according to step 1) in Example 1 with (R)-4-(benzyloxy)-2-fluoro-4-oxobutyric acid as a raw material) was slowly added dropwise. The reaction was monitored by HPLC. After the reaction was completed, the DCM layer was washed with an aqueous hydrochloric acid solution (pH=about 1.0), and the organic layer was dried over anhydrous sodium sulfate and filtered. The solvent was removed by rotary evaporation. The resulting product was dissolved in anhydrous tetrahydrofuran (45 ml), and was hydrogenated in the presence of Pd—C catalyst (5-10%, w/w). After hydrogenation, Pd—C was filtered out, the solvent was removed by evaporation, and the residues were purified by preparative chromatography to obtain a corresponding product. A tert-butanol solution of an equivalent amount of sodium tert-butoxide was slowly added to an anhydrous tetrahydrofuran solution of the resulting product on an ice bath. Solids precipitated out and were filtered, and then the filter cake was washed with a small amount of tetrahydrofuran and dried to obtain compound 11. Mass (ESI) [M-Na]−=435.23.
Epiallopregnanolone (4.5 mmol) and 4-dimethylaminopyridine (8.2 mmol) were dissolved in anhydrous dichloromethane (20 ml) at −40° C., and a dichloromethane solution of (R)-dibenzyl (4-chloro-3-fluoro-4-oxobutyl)phosphate (prepared according to step 1) in Example 1 with (R)-4-((bis(benzyloxy)phosphoryl)oxy)-2-fluorobutyric acid as a raw material) was slowly added dropwise. The reaction was monitored by HPLC. After the reaction was completed, the DCM layer was washed with an aqueous hydrochloric acid solution (pH=about 1.0), and the organic layer was dried over anhydrous sodium sulfate and filtered. The solvent was removed by rotary evaporation. The resulting product was dissolved in anhydrous tetrahydrofuran (45 ml), and was hydrogenated in the presence of Pd—C catalyst (5-10%, w/w). After hydrogenation, Pd—C was filtered out, the solvent was removed by evaporation, and the residues were purified by preparative chromatography to obtain a corresponding acid. A tert-butanol solution of sodium tert-butoxide in an amount equivalent to that of the acid was slowly added to an anhydrous tetrahydrofuran solution of the acid on an ice bath. Solids precipitated out and were filtered, and then the filter cake was washed with a small amount of tetrahydrofuran and dried to obtain compound 16. Mass (ESI) [M-2Na+H]−=501.22.
Epiallopregnanolone (4.5 mmol) and 4-dimethylaminopyridine (8.2 mmol) were dissolved in anhydrous dichloromethane (20 ml) at room temperature, and then dicyclohexyl carbodiimide (10 mmol) and (S)-3-(benzyloxy)-2-fluoro-3-oxopropionic acid (8 mmol) were added. The mixture was allowed to react at room temperature, and the reaction was monitored by HPLC. After the reaction was completed, the DCM layer was washed with an aqueous hydrochloric acid solution (pH=about 1.0), and the organic layer was dried over anhydrous sodium sulfate and filtered. The solvent was removed by rotary evaporation. The resulting product was dissolved in anhydrous tetrahydrofuran (30 ml), and was hydrogenated in the presence of Pd—C catalyst (5-10%, w/w). After hydrogenation, Pd—C was filtered out, the solvent was removed by evaporation, and the residues were purified by preparative chromatography to obtain a corresponding acid. An equivalent amount of arginine was added to an anhydrous tetrahydrofuran solution of the acid on an ice bath. Solids precipitated out and were filtered, and then the filter cake was washed with ice-cold tetrahydrofuran to obtain compound 22. Mass (ESI) [M-ArgH]−=421.22.
The following compounds were prepared according to methods similar to those of Example 1 or Example 2:
Each of the compounds to be tested was weighed (1 mg), placed in a 5% glucose solution (2 ml) and formulated into a solution having a concentration of about 0.5 mg/ml. The solution stood at room temperature, and was sampled at several time points (0.5 h, 1 h, 2 h, 3 h and 5 h). The samples were detected by HPLC, and the results were given below:
The test results show that the dissociation percentages of the tested compounds of the present invention are less than 1% after being placed in a 5% glucose solution (e.g., for 5 h), indicating that the compounds have good stability in a 5% glucose solution and thus are suitable for injection.
It indicates that the compounds provided in the present invention have relatively good solubility in water, thus changing the solubility of epiallopregnanolone.
Each of the compounds to be tested was weighed (1 mg), placed in water (2 ml), and formulated into a solution having a concentration of about 0.5 mg/ml as a stock solution for further use. Whole blood of rats (0.45 ml) was taken and placed in a 2 ml EP tube, which was placed in a 37° C. water bath for preheating for 20 seconds. Then the stock solution of the compound (50 μL) was added quickly. After a homogeneous mixing, the EP tube was placed in a 37° C. water bath, and time counting was initiated. Acetonitrile (1 ml) was injected immediately at predetermined time points (1 min, 3 min, 5 min, 15 min). The solution was centrifuged for 5 min (15000 rpm), filtered with a 0.22 μm filtration membrane and subjected to HPLC detection, and the results were given below:
The test results show that the tested compounds of the present invention can be dissociated in rat plasma to release the active drug (epiallopregnanolone), and the dissociation percents are more than 90% after 15 min, indicating that the epiallopregnanolone derivatives provided in the present invention can rapidly release epiallopregnanolone in the whole blood of rats, thus exerting therapeutic effects.
In addition to those described herein, various modifications to the present invention will be apparent to those skilled in the art based on the foregoing description. Such modifications are intended to fall within the scope of the appended claims. Each reference cited herein (including all patents, patent applications, journal articles, books and any other disclosures) are incorporated herein by reference in its entirety.
| Number | Date | Country | Kind |
|---|---|---|---|
| 201611101726.X | Dec 2016 | CN | national |
This application is a National Stage Entry under 35 U.S.C. § 371 of International Application No. PCT/CN2017/114590, filed Dec. 5, 2017, which claims the benefit of priority from Chinese patent application No. 201611101726.X, filed on Dec. 5, 2016, the contents of which are incorporated herein by reference in their entireties.
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/CN2017/114590 | 12/5/2017 | WO | 00 |
