Patent Application
20220152216
The present invention relates to antibody-polymer conjugates which have high drug carrying capacity and which can realize active targeting thanks to the antibody existing thereon.
The conjugation of humanized or non-humanized monoclonal antibodies by means of chemical bond with cytotoxic small molecule drugs has led to important changes in the design of cancer chemotherapy. (Chari et al, 2014; Perez et al, 2014; Bouchard et al, 2014; Jain et al, 2015; McCombs and Owen, 2015; Chudasama et al., 2016; Diamantis and Banerji, 2016).
The most important advantage provided by antibody drug conjugates is that they provide treatment which is unique to the target and that low cytotoxicity is observed in healthy cells except the target and that low immunogenicity is observed.
In preparation of antibody-drug conjugates, cleavablecleavable bonds like hydrazone, disulphide can be used, or cleavablenon-cleavable bonds like amide can be used.
In the known art, there are antibody-drug conjugates which are sold in the market. As an example to these, Brentuximab vedotin (Adcetris®) can be shown. Adcetris comprises Brentuximab (Mab) which is a monoclonal antibody and mono-methyl auristatin as drug. Said monoclonal antibody and drug molecule are bound to each other by means of Val-Cit linker which can be cleaved in the body by means of Cathepsin B enzyme. In such conjugates, there are on the average 4 drug molecules per an antibody and these are connected to one other by means of a linker cleavable in the body. The bond which exists in the conjugate which reaches the target tumor cell with the help of monoclonal antibody is cleaved here by means of cathepsin B enzyme, and mono-methyl auristatin which is a cyto-toxic agent is released to the tumor region. This way, mono-methyl auristatin molecule, which is not possible to be used in any manner due to high toxicity, can be used for treatment.
Another antibody-drug conjugate sold in the market is Trastuzumab emtansine known with the trade name Kadcyla®. In this product, trastuzumab is used as the antibody, and cytotoxic emtansine, having abbreviation DM1, is used as drug. While trastuzumab binds to the HER2 receptor in a stand-alone manner and stops growth of cancer cells, trastuzumab-emtansine conjugate is carried into the cell by means of receptor and it is catabolized by lysosomes, and by means of this, emtansine is released. The released emtansine is bound to tubulin and stops mitotic cycle and thus, it leads to death of the cancer cell. Since trastuzumab is specific to the HER2 receptor and since HER2 receptor is expressed more than normal only in cancer cells, trastuzumab-emtansine conjugate is specifically transferred to the tumor cells.
In this conjugate, the bond between the drug and the antibody is provided by means of succinimidil trans-4-(maleimidil-methyl)cyclo-hexane-1-carboxylate (SMCC). While the thiol group, which exists on emtansine, reacts with maleimide, the succinimide group which exists on SMCC reacts with lysine which exists on trastuzumab, and amide bond is formed. Approximately 3.5 drug molecules exist for each trastuzumab.
Kopecek et. al., 2018 (Eur. J. Pharm. Sci., 2017 103, 36-46) has developed Epirubicin which is copolymerized with conjugated HPMA by means of Rituximab in order to increase treatment efficiency and patient tolerance in B-cell lymphoma patients. Since the water solubility of the HPMA monomer used in obtaining these conjugates is delimited, a disadvantage occurs.
The antibody-drug conjugates, which are present in the known state of the art, are obtained by means of binding the drug to the antibody by means of a cleavable or non-cleavable bond as in the Adcetris® and Kadcyla® examples given above. In such conjugations, the drug-antibody proportion is in general approximately 4 on the average. In other words, on an antibody, there are drug molecules between 0 and 8, on average there are 4 drug molecules. For the efficiency of the treatment, it is preferred that there are more drug molecules on the antibody, however, when the antibody is modified for providing more drug molecules, the activity of monoclonal antibody decreases, in other words, binding ability decreases.
Moreover, since small amount of drug can be carried in such antibody-drug conjugates, drugs are used which are toxic such that they cannot be given normally in a stand-alone manner. When even a small part of these drugs breaks from the antibody-drug conjugate without reaching the tumor, these drugs lead to serious side effects.
Another problem of the known antibody-drug conjugates occurs during chemical modification of the antibody for binding the linkers or drugs. In order to form an area which is suitable for reaction on the antibody, the disulphide bonds which exist in the antibody structure are broken by means of chemicals named as DTT or TCEP, and the thiol reactive groups are obtained. Since this process leads to a change in the 3-dimensional structure in said antibody, it leads to a decrease in the bonding ability to the related receptor.
The inventors prepared the subject matter antibody-polymer conjugates for increasing the drug amount per antibody and at the same time for developing antibody-polymer conjugates which have higher solubility when compared with the antibody-polymer conjugates which are present in the known art.
The inventors have prepared a drug carrying system (antibody-polymer conjugate) formed by polymer which carries therapeutic agent that has been conjugated with cleavable or non-cleavable bonds to monoclonal antibody and said polymer is characterized by comprising (meth)acrylate backbone and poly(ethylene glycol) at the side branches and the therapeutic agent is connected to the polymer backbone by means of a cleavable linker.
Thanks to the antibody-polymer conjugates according to the invention, the amount of drug per polymer can be changed as desired. Moreover, since the polymers which exist in the antibody-polymer conjugates according to the invention carry more than one drug molecule in each chain, when said polymers are conjugated to an antibody, the drug:antibody proportion is also increased. Thus, the drugs with less cytotoxic effect are directly sent to the tumor tissue by means of the conjugate. Moreover, the poly(ethylene glycol) chains which exist in side branches of the polymer structure provide obtaining antibody-polymer conjugates with high water solubility and thus high bioavailability.
In a preferred embodiment of the present invention, the antibody-polymer conjugates are shown by Formula 1;
wherein;
R1, R2, R3 are independently selected from —H or —CH3
x is a natural number between 1 and 140
y is a natural number between 1 and 40
z is a natural number between 1 and 15
L1 is a cleavable linker
L2 is a linker
D is a therapeutic agent
A is a monoclonal antibody.
The term “antibody-polymer conjugate” used within the scope of the present invention describes the polymer structure connected in a covalent manner to an antibody. The polymeric structures mentioned within the scope of the present invention are polymers having (meth)acrylate backbone and comprising therapeutic agent and poly(ethylene glycol) bound to the polymer backbone by means of covalent bonds at the side chains.
The terms of “antibody-polymer conjugate”, “formula 1”, “antibody-polymer conjugate shown by formula 1” that are used within the scope of the present invention are equivalent to one other and can be used interchangeably.
The term “(meth)acrylate” used within the scope of the present invention indicates “acrylate” and “meth-acrylate” terms. Therefore, “(meth)acrylate” can be used instead of “acrylate” and “methacrylate” and comprises all of the above mentioned characteristics of these terms.
The term “cleavable linker” used within the scope of the present invention describes the bonds which connect the polymer backbone and the therapeutic agent to each other and which can be broken in the presence of a specific stimulant. Here, the stimulant can be any item which can provide breaking of said bond, for instance, light, chemical agent, biologic agent, any substance which can exist in the body of a mammalian, heat etc.
The term “linker” used within the scope of the present invention describes the bonds which connect the polymer backbone and the antibody to each other. Here, the linkers can be cleavable or non-cleavable.
The term “therapeutic agent” used within the scope of the present invention describes all agents used for eliminating, for decelerating, for preventing any physiological condition, for decreasing symptoms or for providing elimination of the symptoms or for detecting the presence of a physiological condition. Said agents can have small molecules, macro-molecules or a biological-based structure or a structure which mimics biological based molecules.
The term “monoclonal antibody” used within the scope of the present invention describes the antibodies which show reaction against only one epitope and which are based on only one B-lymphocyte. The terms “monoclonal antibody” and “antibody” used within the scope of the present invention are equivalent to one other and can be used interchangeably.
The number shown with z provided in the scope of the present invention describes the number of polymer chains connected to an antibody. In a preferred embodiment of the present invention, z describes a number between 1.05 and 14 and preferably a number between 1.5 and 13. In an embodiment of the present invention, z can be a number between 2.0 and 12 or between 2.5 and 11 or between 3 and 10. For instance, z can be a number like 1.05, 1.1, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10.0, 10.5, 11.0.
The drug:antibody ratio which exists in the antibody polymer conjugates of the invention, in other words, the amount of drug molecule per antibody, can be between 1:1 and 100:1. In other words, in the subject matter antibody-polymer conjugates, a drug between 1 and 100 molecules can be provided per antibody. In a preferred embodiment of the present invention, the drug antibody ratio can be between 4:1 and 90:1, and in a particularly embodiment, it can be between 6:1 and 80:1.
The term “poly(ethylene glycol)” used within the scope of the present invention indicates a polyether compound which has the structure of H—(O—CH2—CH2)n-OR3 and n is a natural number between 1 and 200 and R3 is selected from H or —CH3. PEG is defined as an oligomer or polymer of the ethylene oxide. The terms of “PEG”, “polyethylene glycol”, “polyethylene oxide”, “PEO”, “poly-oxy-ethylene” and “POE” indicate the same structure and can be interchangeably used in this specification.
The polymer which has poly(ethylene glycol) and therapeutic agent in the subject matter side chains can be a random copolymer or block copolymer structure. The term “random copolymer” describes a copolymer where the monomers forming the copolymer do not follow any specific sequence. The term “block copolymer” describes a copolymer where all of a kind of monomer are grouped together and where the other type of monomer are grouped together. In a preferred embodiment of the present invention, the polymer having poly(ethylene glycol) and therapeutic agent is in the structure of random copolymer.
In a preferred embodiment of the present invention, in the subject matter antibody-polymer conjugates, the therapeutic agent can be selected from a group comprising anti-neoplastic agents, immune system modulating agents, agents which show anti-angiogenic characteristic on malign cells, agents which inhibit cell proliferation on malign cells or agents which increase immune cell proliferation.
The term “malign cells” used here describes the cells where the cell cycle is rapid and/or where the genetic structure has changed and/or which grow in an uncontrolled manner and/or which have increased mobility and/or where the cell surface structure has changed and/or where lytic factor secretion has changed and/or which show invasive growth and/or which have chemotaxis characteristic and/or which have abnormal shape and/or size and/or where cyto-plasma has decreased and/or which have big core and/or which have grown nucleus pores.
The term “immune cell” used herein describes all of the cells which exist in a mammalian body, preferably the cells which exist in human body. Said cells can show activity in different manners in the immune system. The immune cells described within the scope of the present invention can be B-cells, dendritic cells, granulocytes, natural lymphoid cells, mega-karyocytes, monocytes, macrophages, myeloid-based suppressive cells, platelets, red blood cells, T-cells and thymocytes.
Anti-neoplastic agent can be selected from a sub-group comprising but not limited to nucleoside analogs, antifolates, other metabolites, topoisomerase I inhibitors, anthracyclins, podophyllotoxins, taxanes, vinca alkaloids, alkylating substances, platinum compounds, anti-hormones, radio-pharmaceuticals, tyrosine kinase inhibitors, rapamycin protein complex mammalian target (mTOR) inhibitors, retinoids, immune system regulating substances, histone deacetyase inhibitors and other substances.
Nucleoside analogs can be selected from a group comprising but not limited to azacitidin, cladribin, chlopharabin, citarabin, decitabin, floxuridin, fludarabin, florouracil (5-FU), gemcitabine, mercaptopurine, nelarabin, pentostatin, tioguanin, trifluoridin and tipiracil.
Antifolates can be selected from a group comprising but not limited to metotreksat, pemetrexed, pralatrexed and raltitrexed.
The other metabolites can be selected from a group comprising but not limited to hydroxy-carbamide.
Topoisomerase I inhibitors can be selected from a group comprising but not limited to irinotecan and topotecan.
Anthracyclins can be selected from a group comprising but not limited to daunorubicin, doxorubicin, epirubicin, idarubicin, mitoxantron and valrubicin.
Podophyllotoxins can be selected from a group comprising but not limited to etopocid and tenipocid.
Taxanes can be selected from a group comprising but not limited to cabazitaxel, docetaxel and paclitaxel.
Vinca alkaloids can be selected from a group comprising but not limited to vinblastin, vincristin, vindesin, vinflunin and vinorelbin.
Alkylating substances can be selected from a group comprising but not limited to bendamustin, chlorambucil, dacarbazine, melphalane, streptozotosin and trabestedin.
Antihormon compounds can be selected from a group comprising but not limited to abirateron, bicalutamid, siproteron, degarelix, exemestane, fulvestrant, goserelin, histrelin, leuprolide, miphepristone and triptorelin.
Tyrosine kinase inhibitors can be selected from a group comprising but not limited to afatinib, aksitinib, bosutinib, kobimetinib, crizotinib, dasatinib, erlotinib, gefitinib, imatinib, lapatinib, nilotinib, osimertinib, pazopanib, ruksolitinib, sunitinib and vandetanib.
Rapamycin protein complex mammalian target (mTOR) inhibitors can be selected from a group comprising but not limited to everolimus and temsirolimus.
Retinoids can be selected from a group comprising but not limited to alitretinoin, bexarotene, isotretinoin, tamibarotene and tretinoin.
Immune system regulation substances can be selected from a group comprising but not limited to lenalidomid, pomalidomid and talidomid.
Histone deacetylase inhibitors can be selected from a group comprising but not limited to belinostat, panobinostat, valproate and vorinostat.
The other substances can be selected from a group comprising but not limited to anagrelide, seritinib, dabrafenib, idelalisib, ibrutinib, palbocyclib, vemuraphenib, bleomisin, bortezomib, dactinomisin, eribulin, estramustine, ixabepilone, mitomisin, prokarbazin, alectinib, fluxymesteron, iobenguane, imiguimod, interferon, iksazomib, Ianreotid, lentinan, octreotid, omacetaxine, tegafur, gimerazil, oterasil, uracil, kombretastatin, emtansine and chloroquine.
In a preferred embodiment of the present invention, the therapeutic agent can be selected from antineoplastic agents, preferably from taxanes, antifolates, tyrosine kinase inhibitors, anthracyclins, nucleoside analogs or other substances.
In a most preferred embodiment, the therapeutic agent is selected from a group comprising docetaxel, pemetrexed, hydroxchloroquine, combretastatin, gemcitabine, doxorubicin, 5-Fluorouracil (5-FU), 5′-Deoxy 5-Fluorocytidine (5′-DFCR), emtansine and lapatinib.
In an embodiment of the present invention, the therapeutic agent is docetaxel. In an embodiment of the present invention, the therapeutic agent is hydroxychloroquine. In an embodiment of the present invention, the therapeutic agent is combretastatine. In an embodiment of the present invention, the therapeutic agent is gemcitabine. In an embodiment of the present invention, the therapeutic agent is doxorubicin. In an embodiment of the present invention, the therapeutic agent is 5-FU. In an embodiment of the present invention, the therapeutic agent is 5′-deoxy 5-fluorocytidine (5′-DFCR). In an embodiment of the present invention, the therapeutic agent is lapatinib. In an embodiment of the present invention, the therapeutic agent is emtansine.
In an embodiment of the present invention, the therapeutic agent is a combination of two or more therapeutic agents selected from the group comprising docetaxel, pemetrexed, chloroquine, combretastatine, gemcitabine, doxorubicin, 5-FU, 5′-Deoxy 5-Fluorocytidine (5′-DFCR), emtansine and lapatinib.
The therapeutic agent can be present at an amount between 1% and 40% of the polymer weight provided on the antibody-polymer conjugate, preferably at an amount between 2% and 35% and particularly preferably between 3% and 30%. The therapeutic agent can be present at an amount between 4% and 25% or between 5% and 24% or between 6% and 23% or between 7% and 22% or between 8% and 20% of the polymer weight which exists on the antibody-polymer conjugate.
Any monoclonal antibody can be used as antibody in antibody-polymer conjugates prepared within the scope of the present invention. In a preferred embodiment of the present invention, the monoclonal antibody can be selected from a group comprising 3F8, 8H9, Abagovomab, Abciximab, Abituzumab, Abrezekimab, Abrilumab, Actoxumab, Adalimumab, Adecatumumab, Atidortoxumab, Aducanumab, Afasevikumab, Afelimomab, Alacizumab pegol, Alemtuzumab, Alirocumab, Altumomab pentetate, Amatuximab, Anatumomab mafenatox, Andecaliximab, Anetumab ravtansine, Anifrolumab, Anrukinzumab (IMA-638), Apolizumab, Aprutumab ixadotin, Arcitumomab, Ascrinvacumab, Aselizumab, Atezolizumab, Atinumab, Atorolimumab, Avelumab, Azintuxizumab vedotin, Bapineuzumab, Basiliximab, Bavituximab, bCD-100, Bectumomab, Begelomab, Belantamab mafodotin, Belimumab, Bemarituzumab, Benralizumab, Berlimatoxumab, Bermekimab, Bersanlimab, Bertilimumab, Besilesomab, Bevacizumab, Bezlotoxumab, Biciromab, Bimagrumab, Bimekizumab, Birtamimab, Bivatuzumab mertansine, Bleselumab, Blinatumomab, Blontuvetmab, Blosozumab, Bococizumab, Brazikumab, Brentuximab vedotin, Briakinumab, Brodalumab, Brolucizumab, Brontictuzumab, Burosumab, Cabiralizumab, Camidanlumab tesirine, Camrelizumab, Canakinumab, Cantuzumab mertansine, Cantuzumab ravtansine, Caplacizumab, Capromab pendetide, Carlumab, Carotuximab, Catumaxomab, CBR96-doxorubicin immunoconjugate, Cedelizumab, Cemiplimab, Cergutuzumab amunaleukin, Certolizumab pegol, Cetrelimab, Cetuximab, Cibisatamab, Cirmtuzumab, Citatuzumab bogatox, Cixutumumab, Clazakizumab, Clenoliximab, Clivatuzumab tetraxetan, Codrituzumab, Cofetuzumab pelidotin, Coltuximab ravtansine, Conatumumab, Concizumab, Cosfroviximab, Crenezumab, Crizanlizumab, Crotedumab, CR6261, Cusatuzumab, Dacetuzumab, Daclizumab, Dalotuzumab, Dapirolizumab pegol, Daratumumab, Dectrekumab, Demcizumab, Denintuzumab mafodotin, Denosumab, Depatuxizumab mafodotin, Derlotuximab biotin, Detumomab, Dezamizumab, Dinutuximab, Diridavumab, Domagrozumab, Dorlimomab aritox, Dostarlimab, Drozitumab, DS-8201, Duligotuzumab, Dupilumab, Durvalumab, Dusigitumab, Duvortuxizumab, Ecromeximab, Eculizumab, Edobacomab, Edrecolomab, Efalizumab, Efungumab, Eldelumab, Elezanumab, Elgemtumab, Elotuzumab, Elsilimomab, Emactuzumab, Emapalumab, Emibetuzumab, Emicizumab, Enapotamab vedotin, Enavatuzumab, Enfortumab vedotin, Enlimomab pegol, Enoblituzumab, Enokizumab, Enoticumab, Ensituximab, Epitumomab cituxetan, Epratuzumab, Eptinezumab, Erenumab, Erlizumab, Ertumaxomab, Etaracizumab, Etigilimab, Etrolizumab, Evinacumab, Evolocumab, Exbivirumab, Fanolesomab, Faralimomab, Faricimab, Farletuzumab, Fasinumab, FBTA05, Felvizumab, Fezakinumab, Fibatuzumab, Ficlatuzumab, Figitumumab, Firivumab, Flanvotumab, Fletikumab, Flotetuzumab, Fontolizumab, Foralumab, Foravirumab, Fremanezumab, Fresolimumab, Frovocimab, Frunevetmab, Fulranumab, Futuximab, Galcanezumab, Galiximab, Gancotamab, Ganitumab, Gantenerumab, Gatipotuzumab, Gavilimomab, Gedivumab, Gemtuzumab ozogamicin, Gevokizumab, Gilvetmab, Gimsilumab, Girentuximab, Glembatumumab vedotin, Golimumab, Gomiliximab, Gosuranemab, Guselkumab, lanalumab, Ibalizumab, 1B1308, Ibritumomab tiuxetan, Icrucumab, Idarucizumab, Ifabotuzumab, Igovomab, Iladatuzumab vedotin, IMAB362, Imalumab, Imaprelimab, Imciromab, Imgatuzumab, Inclacumab, Indatuximab ravtansine, Indusatumab vedotin, Inebilizumab, Infliximab, Intetumumab, Inolimomab, Inotuzumab ozogamicin, Ipilimumab, Iomab-B, Iratumumab, Isatuximab, Iscalimab, Istiratumab, Itolizumab, Ixekizumab, Keliximab, Labetuzumab, Lacnotuzumab, Ladiratuzumab vedotin, Lampalizumab, Lanadelumab, Landogrozumab, Laprituximab emtansine, Llarcaviximab, Lebrikizumab, Lemalesomab, Lendalizumab, Lenvervimab, Lenzilumab, Lerdelimumab, Leronlimab, Lesofavumab, Letolizumab, Lexatumumab, Libivirumab, Lifastuzumab vedotin, Ligelizumab, Loncastuximab tesirine, Losatuxizumab vedotin, Lilotomab satetraxetan, Lintuzumab, Lirilumab, Lodelcizumab, Lokivetmab, Lorvotuzumab mertansine, Lucatumumab, Lulizumab pegol, Lumiliximab, Lumretuzumab, Lupartumab amadotin, Lutikizumab, Mapatumumab, Margetuximab, Marstacimab, Maslimomab, Mavrilimumab, Matuzumab, Mepolizumab, Metelimumab, Milatuzumab, Minretumomab, Mirikizumab, Mirvetuximab soravtansine, Mitumomab, Modotuximab, Mogamulizumab, Monalizumab, Morolimumab, Mosunetuzumab, Motavizumab, Moxetumomab pasudotox, Muromonab-CD3, Nacolomab tafenatox, Namilumab, Naptumomab estafenatox, Naratuximab emtansine, Narnatumab, Natalizumab, Navicixizumab, Navivumab, Naxitamab, Nebacumab, Necitumumab, Nemolizumab, NEOD001, Nerelimomab, Nesvacumab, Netakimab, Nimotuzumab, Nirsevimab, Nivolumab, Nofetumomab merpentan, Obiltoxaximab, Obinutuzumab, Ocaratuzumab, Ocrelizumab, Odulimomab, Ofatumumab, Olaratumab, Oleclumab, Olendalizumab, Olokizumab, Omalizumab, Omburtamab, OMS721, Onartuzumab, Ontuxizumab, Onvatilimab, Opicinumab, Oportuzumab monatox, Oregovomab, Orticumab, Otelixizumab, Otilimab, Otlertuzumab, Oxelumab, Ozanezumab, Ozoralizumab, Pagibaximab, Palivizumab, Pamrevlumab, Panitumumab, Pankomab, Panobacumab, Parsatuzumab, Pascolizumab, Pasotuxizumab, Pateclizumab, Patritumab, PDR001, Pembrolizumab, Pemtumomab, Perakizumab, Pertuzumab, Pexelizumab, Pidilizumab, Pinatuzumab vedotin, Pintumomab, Placulumab, Plozalizumab, Pogalizumab, Polatuzumab vedotin, Ponezumab, Porgaviximab, Prasinezumab, Prezalizumab, Priliximab, Pritoxaximab, Pritumumab, PRO 140,Quilizumab, Racotumomab, Radretumab, Rafivirumab, Ralpancizumab, Ramucirumab, Ranevetmab, Ranibizumab, Raxibacumab, Ravagalimab, Ravulizumab, Refanezumab, Regavirumab, Relatlimab, Remtolumab, Reslizumab, Rilotumumab, Rinucumab, Risankizumab, Rituximab, Rivabazumab pegol, Robatumumab, Rmab, Roledumab, Romilkimab, Romosozumab, Rontalizumab, Rosmantuzumab, Rovalpituzumab tesirine, Rovelizumab, Rozanolixizumab, Ruplizumab, SA237, Sacituzumab govitecan, Samalizumab, Samrotamab vedotin, Sarilumab, Satralizumab, Satumomab pendetide, Secukinumab, Selicrelumab, Seribantumab, Setoxaximab, Setrusumab, Sevirumab, Sibrotuzumab, SGN-CD19A, SHP647, Sifalimumab, Siltuximab, Simtuzumab, Siplizumab, Sirtratumab vedotin, Sirukumab, Sofituzumab vedotin, Solanezumab, Solitomab, Sonepcizumab, Sontuzumab, Spartalizumab, Stamulumab, Sulesomab, Suptavumab, Sutimlimab, Suvizumab, Suvratoxumab, Tabalumab, Tacatuzumab tetraxetan, Tadocizumab, Talacotuzumab, Talizumab, Tamtuvetmab, Tanezumab, Taplitumomab paptox, Tarextumab, Tavolimab, Tefibazumab, Telimomab aritox, Telisotuzumab vedotin, Tenatumomab, Teneliximab, Teplizumab, Tepoditamab, Teprotumumab, Tesidolumab, Tetulomab, Tezepelumab, TGN1412, Tibulizumab, Tildrakizumab, Tigatuzumab, Timigutuzumab, Timolumab, Tiragotumab, Tislelizumab, Tisotumab vedotin, TNX-650, Tocilizumab, Tomuzotuximab, Toralizumab, Tosatoxumab, Tositumomab, Tovetumab, Tralokinumab, Trastuzumab, Trastuzumab emtansine, TRBSO7, Tregalizumab, Tremelimumab, Trevogrumab, Tucotuzumab celmoleukin, Tuvirumab, Ublituximab, Ulocuplumab, Urelumab, Urtoxazumab, Ustekinumab, Utomilumab, Vadastuximab talirine, Vanalimab, Vandortuzumab vedotin, Vantictumab, Vanucizumab, Vapaliximab, Varisacumab, Varlilumab, Vatelizumab, Vedolizumab, Veltuzumab, Vepalimomab, Vesencumab, Visilizumab, Vobarilizumab, Volociximab, Vonlerolizumab, Vopratelimab, Vorsetuzumab mafodotin, Votumumab, Vunakizumab, Xentuzumab, XMAB-5574, Zalutumumab, Zanolimumab, Zatuximab, Zenocutuzumab, Ziralimumab Zolbetuximab (=IMAB362, Claudiximab), Zolimomab aritox. In a preferred embodiment of the present invention, an antibody selected from a group comprising nivolulumab, pembrolizumab trastuzumab, cetuximab is used. In a particularly preferred embodiment of the present invention, trastuzumab is used.
The linker (L2) used in the antibody-polymer conjugates according to the invention can be selected from cleavable linkers (L1) or non-cleavable linkers as also mentioned in the definition given above. Said non-cleavable linkers describe the bonds which protect their forms under the physiologic conditions of the body and which do not fragment or which fragment in negligible amounts. The non-cleavable linker can be formed by C1-C10 hydrocarbon chain and optionally one or more carbons, which exist(s) in said hydrocarbon chain can be substituted with —O, —NH, —N, —S, —C(O) or with the combinations thereof and said carbons can be adjacent to or separate from each other. In a preferred embodiment of the present invention, linkers according to the invention can comprise one or more substitution like —COOH, —OH, —NH2, —SH— in addition to —O, —NH, —N, —S, —C(O) substitutions or in an independent manner from —O, —NH, —N, —S, —C(O) substitutions. In a particularly preferred embodiment of the present invention, as the linker (L2), —C(O)CH2CH2C(CH3)(CN)— is used, wherein the carbonyl group shown by —C(O) reacts with —NH2 provided on any amino acid provided in monoclonal antibody structure and is connected to the amino acid by means of amide bond.
The cleavable linker (L1) used in the subject matter antibody-polymer conjugates can be any substance like poly(ethylene glycol), an amino acid, poly(amino acid) (for instance, peptide or oligo-peptide) or poly(peptide). In a preferred embodiment of the present invention, the cleavable linker can be selected from short peptides which have specific peptide sequences such as Gly-Phe-Leu-Gly which can be described as GFLG or Val-Cit or Phe-Lys or Val-Ala or Ala-Leu-Ala-Leu which can be broken by catepcine B.
The cleavable linker (L1) used in the subject matter antibody-polymer conjugates can be C1-C10 hydrocarbon which is substituted with at least one acetal, ester, imine, amide, disulphide, carbamate or with binary or ternary combinations thereof.
In another embodiment of the present invention, the cleavable linker (L1) can be the combination of a peptide chain selected from a group comprising Phe-Lys or Val-Ala or Ala-Leu-Ala-Leu or Val-Cit, GFLG and a substituted C1-C10 hydrocarbon comprising at least one functional group selected from a group comprising acetal, ester, imine, amide, disulphide, carbonate, carbamate, hydrazone.
In the case, where L2 is selected from cleavable linkers (L1), the L1 binding the drug moiety to the polymer backbone and L2 which is binding the polymer drug conjugate to the antibody are independent from each other and can be the same or different from another as can be seen in the below embodiments of the invention.
In a preferred embodiment of the present invention, the subject matter antibody-polymer conjugate can comprise one or a number of the below mentioned,
In another aspect, a preferred embodiment of the present invention relates to a method (Method 1) for use in preparing antibody-polymer conjugates shown by Formula 1 wherein said method comprises the following steps:
wherein R1, R2, R3 are independently selected from —H or —CH3, L1 is a cleavable bond, D is a therapeutic agent and n is a natural number between 1 and 200,
In another aspect in a preferred embodiment of the present invention, the present invention relates to a method (Method 2) for use in preparing antibody-polymer conjugates shown by Formula 1, wherein said method comprises the following steps:
In method 1 and method 2, in step a) another functional group like —CN, —N3 is optionally provided on trithiocarbonate or dithioate molecule where carboxylic acid functional group is provided thereon. Said trithiocarbonate or dithioate molecule, whereon carboxylic acid functional group is provided, can have any structure provided that there is a trithiocarbonate or dithioate group and at least one carboxylic acid group.
In method 1 and method 2, in the polymerization realized in step b), a second polymerization initiator can be used which is present in the known state of the art. In a preferred embodiment of the present invention, as the second polymerization initiator, 2,2′-Azobis(2-methyl-propionitril) (AIBN) or derivatives, for instance, 4,4′-Azobis(4-cyano-valeric acid), 4,4′-Azobis(4-cyano-valeric acid), 1,1′-Azobis(cyclo-hexane-carbo-nitrile), 2,2′-Azobis(2-metil-propionamidine) dihydro-chloride can be used.
In method 1 and method 2, the polymerization described in step b) is realized at a temperature between 25° C. and 100° C. and preferably at a temperature between 60° C. and 90° C.
In method 1 and method 2, the antibody-polymer conjugation is realized at a temperature between 20° C. and 45° C. and preferably at a temperature of room temperature, particularly preferably at a temperature of 25° C.
In an embodiment of the present invention, PEG (meth)acrylate (Formula II) is used which has an average molecular weight between 200 and 2000 grams/mole. In a preferred embodiment, PEG (meth)acrylate (Formula II) has an average molecular weight between 250 and 1500 grams/mole and in a more preferred embodiment, PEG (meth)acrylate (Formula II) has an average molecular weight between 300 and 1100 grams/mole. The subject matter PEG (meth)acrylate (Formula II) can have an average molecular weight of for instance 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1050, 1100, 1150, 1200, 1250, 1300, 1350, 1400, 1450, 1500, 1550, 1600, 1650, 1700, 1750, 1800, 1850, 1900, 1950, 2000 grams/mole.
Monoclonal antibodies provided in antibody-polymer conjugates provide entry of the attached therapeutic agents into the cell when they go to the target region with the help of their affinity towards the highly expressed receptors in the cells. Thus, the targeted cell type changes according to the used mAb type. The subject matter antibody-polymer conjugates can be used in the treatment of different diseases according to the characteristics of the monoclonal antibody which forms the conjugate.
As used herein, the terms of “treatment” or “treating” describes elimination or reduction of at least one of biological markers (for instance, tumor or tumor markers) which show the presence of a disease or prevention, reduction, mitigation, healing or blocking of at least one symptom which characterizes a pathological disease in a subject where a disease diagnosis has been realized or who is determined to have high probability of having a disease.
The subject matter antibody-polymer conjugates in general can be applied in the form of a pharmaceutical composition.
Therefore, an embodiment of the present invention is pharmaceutical compositions comprising antibody-polymer conjugates shown by formula I and at least one pharmaceutically acceptable excipient.
The composition comprising the subject matter antibody-polymer conjugate can be at any suitable form based on the preferred method in terms of application of this composition to a patient. The composition comprising the subject matter antibody-polymer conjugates can be formulated in a manner applied orally for instance in the form of liquid dispersions or aqueous or oily suspensions or the composition can be formulated for parenteral application like for instance, subcutaneous, intravenous, intramuscular, intrasternal, intra-peritoneal, intradermal, transdermal or other infusion methods. The subject matter composition comprising polymer drug conjugates can also be formulated for application by means of respiratory route in the form of spray tube or for application as solution for application by means of respiratory device or nebulizer. The subject matter antibody-polymer conjugate is preferably applied transdermally, subcutaneously, intranasally, intravenously, intramuscularly, intratumorally. In any case, the most suitable form in terms of application will depend on the specific therapeutic agent provided in the subject matter antibody-polymer conjugate, the kind of the disease and the physical condition of the subject.
The subject matter antibody-polymer conjugates can be applied together for instance, simultaneously, sequentially or separately with one or more therapeutic agent which can be in the form of a cancer preventing substance or immune system regulating substance, antiviral, anti-infective, anti-microbial or anaesthetic substance or the combinations thereof.
Said second therapeutic agent can be any therapeutic agent provided that it is different from the substance which is present in the antibody-polymer conjugate of the invention.
Within the scope of this specification, it is aimed that the term comprising describes the term including.
In places where it is technically suitable, the embodiments of the invention can be combined.
The embodiments are explained here so as to comprise specific characteristics/elements. The description moreover covers the other embodiments comprising said characteristics/elements or formed by these.
Patents, applications, scientific publications and similar technical references have been included in this document by means of reference.
The embodiments which are described here in a specific manner and clearly described can form the base of a disclaimer in a stand-alone manner together with one or a number of other embodiments.
Now the present invention will be explained with reference to the examples given below which are only for exemplary purposes and which shall not be interpreted as delimiting the scope of the present invention in any manner.
2-(pyridine-2-yl sulphonyl) ethanol (41.7 mg, 0.223 mmol) is dissolved in 4-cyano-4-(dodesyl sulphanyl thio-carbonyl) sulphanyl pentanoic acid (60 mg, 0.148 mmol) and 4-dimethyl-amino-pyridine (DMAP) (18 mg, 0.147 mmol) CH2Cl2 (2 mL). The reaction mixture is cooled to 0° C. in an ice bath, and N,N-dicyclohexyl-carbodiimide (DCC) (36.8 mg, 0.178 mmol) dissolved in CH2Cl2 (2 mL) is added to the reaction mixture dropwise. The reaction is stirred at 25° C. The formed dicyclohexyl urea (DCU) is filtered and removed. 1H NMR (CDCl3, δ, ppm) 8.49 (d, 1H, J=3.8 Hz), 7.67-7.64 (m, 2H), 7.12 (bs, 1H), 4.38 (t, 2H, J=6.3 Hz), 3.33 (t, 2H, J=7.4 Hz), 3.05 (t, 2H, J=6.3 Hz), 2.63-2.33 (m, 4H), 1.87 (s, 3H), 1.68 (dd, 2H, J=14.5, 7.1), 1.39-1.26 (m, 18H), 0.88 (t, 3H, J=6.4 Hz).
4-cyano-4-(phenyl-carbono-thioyl-thio) pentanoic acid (CPADB) (200 mg, 0.72 mmol) is dissolved in N-hydroxy succinimide (NHS) (91 mg, 0.79 mmol) CH2Cl2 (1.5 mL). N,N-Dicyclohexylcarbodimide (DCC) (163 mg, 0.79 mmol) is dissolved in CH2Cl2 (0.5 mL) and is added dropwise to the reaction mixture in the ice bath. The reaction is stirred for 3 hours at room temperature. The obtained raw product is precipitated in cold ethyl acetate and purified by means of product column chromatography. 1H NMR (400 MHz, CDCl3, δ, ppm) δ 7.93 (d, J=7.4 Hz, 2H), 7.58 (t, J=7.4 Hz, 1H), 7.41 (t, J=7.8 Hz, 2H), 3.03-2.95 (m, 2H), 2.86 (s, 4H), 2.79-2.69 (m, 1H), 2.61-2.52 (m, 1H), 1.96 (s, 3H).
Polyethylene glycol monomethyl methacrylate (PEGMEMA) (100 mg, 0.33 mmol) is dissolved in DMF (0.3 mL). PDS-CDTPA (6.8 mg, 0.01 mmol) is dissolved in DMF (0.1 mL) and AIBN (0.39 mg, 2.4×10−3 mmol) is dissolved in DMF (0.1 mL). The prepared solutions are added into PEGMEMA solution under N2 atmosphere. MA-GFLG-DTX (45 mg, 0.04 mmol) solution, prepared by dissolving in DMF (0.4 mL) beforehand is added to the reaction mixture. The reaction is stirred at temperature of 80° C. DMF is removed after the reaction.
The obtained product is precipitated in ether, and the formed precipitate is removed by filtration and purified by means of dialysis membrane. The molecular weight of the obtained polymer is detected by means of GPC (10 kDa). From the 1H NMR result, it has been detected that there is drug with proportion of 10.6% by weight in the polymer chain.
PEGMEMA (86 mg, 0.29 mmol), MA-GFLG-DTX (120 mg, 0.096 mmol) and SCPDB (3.6 mg, 0.096 mmol) are dissolved in DMF (1.00 mL). AIBN (0.36 mg, 0.0019 mmol) is added into the reaction mixture. DMF is removed after polymerization. The obtained raw product is purified by means of dialysis. The molecular weight of the obtained polymer is detected by means of GPC (10 kDa). From the 1H NMR result, it has been detected that there is drug with proportion of 12% by weight in the polymer chain.
2-Imino-thiolane (0.1 mg, 3.6×10−3 mmol) is dissolved in PBS (3 mL, pH 8.2). Trastuzumab (Tmab) (4.6 mg/mL, 3.1×10−5 mmol) is added to the reaction. The reaction is stirred for 2.5 hours at 25° C. The number of thiol groups attached to Tmab is determined by means of Ellman procedure (1). As a result of this experiment, it has been seen that 6 thiols are obtained on each Tmab. Dialysis is made with PBS buffer solution (pH 6.5). In order to determine Tmab concentration, BCA analysis is made (2). As a result of the analysis, it has been detected that Tmab concentration is 3.7 mg/mL.
Tmab-SH and PDS-POEGMEMA-DTX Copolymer is dissolved in DMSO and PBS buffer solution (pH 6.5). The obtained mixture is stirred at room temperature. The obtained product is purified by means of conventional methods.
Tmab and NHS-POEGMEMA-DTX copolymer is dissolved in DMSO and PBS buffer solution (pH 6.5). The obtained mixture is stirred at room temperature. The obtained product is purified by means of conventional methods.
| Number | Date | Country | Kind |
|---|---|---|---|
| 2019/11542 | Jul 2019 | TR | national |
This application is a continuation of PCT International Application No. PCT/TR2020/050674, filed Jul. 29, 2020, which claims priority to Turkish Application No. 2019/11542, filed Jul. 30, 2019, which are hereby incorporated by reference in its entirety.
| Number | Date | Country | |
|---|---|---|---|
| Parent | PCT/TR2020/050674 | Jul 2020 | US |
| Child | 17587667 | US |
