CN117731798A - Antibody coupling drug, intermediate thereof, preparation method and application - Google Patents
Antibody coupling drug, intermediate thereof, preparation method and application Download PDFInfo
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- CN117731798A CN117731798A CN202311489217.9A CN202311489217A CN117731798A CN 117731798 A CN117731798 A CN 117731798A CN 202311489217 A CN202311489217 A CN 202311489217A CN 117731798 A CN117731798 A CN 117731798A
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- 238000000926 separation method Methods 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 239000010703 silicon Substances 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical class O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 description 1
- 238000013112 stability test Methods 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 201000011549 stomach cancer Diseases 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
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- 239000000375 suspending agent Substances 0.000 description 1
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- A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
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- A61K47/6873—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site the antibody targeting an immunoglobulin; the antibody being an anti-idiotypic antibody
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- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
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- A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
- A61K47/65—Peptidic linkers, binders or spacers, e.g. peptidic enzyme-labile linkers
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- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
- A61K47/6801—Drug-antibody or immunoglobulin conjugates defined by the pharmacologically or therapeutically active agent
- A61K47/6803—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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Abstract
The invention discloses an antibody coupling drug, an intermediate thereof, a preparation method and application. The invention discloses an antibody coupling drug, the structural general formula of which is Ab- (L) 3 ‑L 2 ‑L 1 ‑D) m . The antibody coupling medicine can realize the wide application of cytotoxic medicine, especially camptothecine in ADC field, and treat tumor patient with microtubule ADC medicine resistance.
Description
The patent application is a divisional application of a patent application with the application number of 201910576227.3, and the application date of the patent application is 2019, 6 and 28, and the invention is named as an antibody coupling drug, an intermediate, a preparation method and application.
Technical Field
The invention belongs to the field of biotechnology and medicine, and particularly relates to an antibody coupling drug, an intermediate thereof, a preparation method and application.
Background
Antibody-conjugated drugs (ADCs) are one of the hot spots of recent interest in the pharmaceutical industry. Because of the inadequate clinical efficacy of many antibody drugs, many industries are increasingly turning their eyes to ADC drugs. Currently, four ADC drugs are commercially available in the world. The FDA approved for the use of dent company Gemtuzumab Ozogamicin (trade name Mylotarg) at 5.17.2000 for the treatment of Acute Myelogenous Leukemia (AML) patients with first relapse, over 60 years old, cd33+, unsuitable for cytotoxic chemotherapy, although the drug was withdrawn in 2010 but re-marketed in 2017, the same year as dent Inotuzumab ozogamicin (trade name Besponsa) was also FDA-approved for the use of refractory B-cell ALL in adult relapse. On month 8 and 19 2011, brentuximab Vedotin (trade name, addetris) developed by Seattle Genetics company was approved by the FDA for the treatment of CD30 positive Hodgkin Lymphoma (HL) and rare disease Systemic Anaplastic Large Cell Lymphoma (SALCL). 22.2013, ado-trastuzumab emtansine (T-DM 1, trade name Kadcyla) developed by Genntech corporation was approved by the FDA for sale, and was used mainly for the treatment of Her2 positive advanced (metastatic) breast cancer. In addition, more than 100 ADC drugs are internationally in clinical and preclinical development stages.
The linker of ADC is a vital component, and its stability not only affects the release of small molecules in tumor cells, but also is related to toxic side effects of ADC. An over-stable linker is generally not readily releasable, such as T-DM1; too labile linkers tend to release the drug in the plasma, thereby bringing about toxicity, such as IMMU-132. The first co-company published patent (patent number ZL 201380053256) discloses a connecting group which is quite stable in blood plasma, namely, a camptothecine compound is connected with an antibody through a section of tetrapeptide which can be digested and cleaved by self-cleaving aminomethylether structure, so that the camptothecine compound has a better anti-tumor effect. Although this structure maintains good stability in plasma, the aminomethyl ether structure is acid-labile, and there are many acidic environments in the body, including both part of the tumor tissue and some normal tissue. Thus ADC releases part of the drug molecule without endocytosis by tumor cells, resulting in toxicity.
The present invention is directed to solving the above-mentioned problems of the prior art. The invention provides two technical schemes, one is to replace an aminomethyl ether structure with a carbon amide structure, so that the stability is enhanced. The other is to directly connect the self-cleavage fragments to the drug molecules through alkyl groups, and the structure has good stability and can generate rapid 1, 6-elimination after enzyme digestion so as to release the drug. Therefore, the invention can solve the defects of the ADC, can keep better biological activity, and can obtain better anti-tumor activity and lower toxicity in vivo.
Disclosure of Invention
The invention aims to overcome the defect of single type of the existing antibody coupling drug, and provides an antibody coupling drug, an intermediate, a preparation method and application. The antibody coupling medicine can realize the wide application of the cytotoxicity medicine in the ADC field and treat tumor patients with microtubule ADC drug resistance.
The invention solves the technical problems through the following technical proposal.
The invention provides an antibody coupling drug, the structural general formula of which is Ab- (L) 3 -L 2 -L 1 -D) m ；
Wherein Ab is an antibody;
d is a cytotoxic drug;
m is 2-8;
L 1 the structure of (C) is shown as a formula I or II; wherein the a-terminus is linked to said cytotoxic agent, the e-terminus is linked to said L 2 Is connected with the c end of the (C) chain,
l is independently a phenylalanine residue, glycine residue, glutamic acid residue, aspartic acid residue, cysteine residue, glutamic acid residue, histidine residue, isoleucine residue, leucine residue, lysine residue, methionine residue, proline residue, serine residue, threonine residue, tryptophan residue, tyrosine residue or valine residue; p is 0 to 4;
R 1 is hydrogen, substituted or unsubstituted C 1 ～C 10 Alkyl, substituted or unsubstituted C 6 ～C 14 Aryl, substituted or unsubstituted 5-to 10-membered heterocyclic group, or substituted or unsubstituted phenoxy; the substituted C 1 ～C 10 Alkyl, substituted C 6 ～C 14 The substituents in the aryl group, the substituted 5-to 10-membered heterocyclic group, and the substituted phenoxy group are one or more groups selected from the group consisting of, when a plurality of substituents are present, the substituents are the same or different: halogen, hydroxy, -NR 1-1 R 1-2 、-S(O) 2 R 1-3 、C 1 ～C 4 Alkyl, C 1 ～C 4 Alkoxy, hydroxy-substituted C 1 ～C 4 Alkoxy, C 3 ～C 8 Cycloalkyl, 5-8 membered heterocyclyl, C 6 ～C 10 Aryl and 5-to 10-membered heteroaryl; the heteroatom in the substituted or unsubstituted 5-10 membered heterocyclic group, 5-8 membered heterocyclic group or 5-10 membered heteroaryl group is selected from one or more of N, O and S, and the number of the heteroatom is 1, 2, 3 or 4; r is R 1-1 、R 1-2 And R is 1-3 Independently C 1 ～C 4 An alkyl group;
R 2 independently hydrogen, substituted or unsubstituted C 1 ～C 10 Alkyl, substituted or unsubstituted C 3 ～C 10 Cycloalkyl, substituted or unsubstituted C 6 ～C 14 Aryl, substituted or unsubstituted 5-10 membered heteroaryl; said substituted C 1 ～C 10 Alkyl, substituted C 3 ～C 10 Cycloalkyl, substituted C 6 ～C 14 The substituents in aryl, and substituted 5-to 10-membered heteroaryl groups are one or more groups selected from the group consisting of, when multiple substituents are present, the substituents are the same or different: halogen, hydroxy, C 1 ～C 4 Alkyl, C 1 ～C 4 Alkoxy, C 3 ～C 8 Cycloalkyl, 5-8 membered heterocyclyl, C 6 ～C 10 Aryl and 5-to 10-membered heteroaryl; the hetero atoms in the 5-8 membered heterocyclic group, the 5-10 membered heteroaryl group and the substituted or unsubstituted 5-10 membered heterocyclic group are selected from one or more of N, O and S, and the number of the hetero atoms is 1, 2, 3 or 4;
n 1 2, 3 or 4;
l' is independently a phenylalanine residue, glycine residue, glutamic acid residue, aspartic acid residue, cysteine residue, glutamic acid residue, histidine residue, isoleucine residue, leucine residue, lysine residue, methionine residue, proline residue, serine residue, threonine residue, tryptophan residue, tyrosine residue or valine residue; p' is 2-4;
L 2 is that Wherein, the c end and L 1 Is connected to the e terminal, the f terminal and the L terminal 3 Is connected with d end of n 2 Independently 1 to 8;
L 3 is thatWherein the b-terminus is linked to said Ab, the d-terminus is linked to said L 2 Is connected to the f terminal of (c).
In a preferred embodiment of the invention, the antibody may be an antibody conventional in the field of anti-tumor ADCs, preferably Trastuzumab or a variant thereof, anti-B7-H3 antibody P2E5 or a variant thereof, anti-claudin 18.2 antibody IMAB362 or a variant thereof, or anti-Trop 2 antibody RS7 or a variant thereof, more preferably Trastuzumab or a variant thereof, most preferably Trastuzumab. The amino acid sequence of the light chain in the anti-HER 2 antibody Trastuzumab is preferably shown as SEQ ID No.5 in a sequence table, and the amino acid sequence of the heavy chain is preferably shown as SEQ ID No.6 in the sequence table. The amino acid sequence of the light chain in the anti-B7-H3 antibody P2E5 is preferably shown as SEQ ID No.7 in a sequence table, and the amino acid sequence of the heavy chain is preferably shown as SEQ ID No.8 in the sequence table. The amino acid sequence of the light chain in the Claudin18.2 antibody IMAB362 is preferably shown as SEQ ID No.1 in a sequence table, and the amino acid sequence of the heavy chain is preferably shown as SEQ ID No.2 in the sequence table. The amino acid sequence of the light chain in the anti-Trop 2 antibody RS7 is preferably shown as SEQ ID No.3 in the sequence table, and the amino acid sequence of the heavy chain is preferably shown as SEQ ID No.4 in the sequence table.
In a preferred embodiment of the present invention, said L 3 Preferably, the b-segment of (2) is linked to the thiol group of said antibody in the form of a thioether bond. To be used forFor example, a->With cysteine residues in said antibodiesThe connection form of the base is->
In a preferred embodiment of the present invention, D may be a cytotoxic drug conventional in the ADC field, and the present invention is particularly preferably a hydroxyl-containing or amino-containing cytotoxic drug, more preferably a hydroxyl-containing or amino-containing topoisomerase inhibitor, still more preferably a hydroxyl-containing or amino-containing topoisomerase I inhibitor, still more preferably camptothecin or a derivative thereof, and most preferablyThe L is 1 Preferably linked to the hydroxyl group in the cytotoxic drug in the form of an ether linkage or to the amino group in the cytotoxic drug. When said L 1 When linked to an amino group in said cytotoxic drug, the hydrogen on said amino group is not substituted or is replaced by an R 3 Substitution; said R is 3 Is C 1 ～C 6 Alkyl, C 3 ～C 8 Cycloalkyl, C 6 ～C 14 Aryl, 5-to 10-membered heteroaryl or-C (=o) R 3-1 The method comprises the steps of carrying out a first treatment on the surface of the The hetero atom in the 5-10 membered heteroaryl is selected from one or more of N, O and S, and the number of the hetero atom is 1, 2, 3 or 4; r is R 3-1 C substituted by hydroxy 1 ～C 4 An alkyl group. When said L 1 After being connected with the D, L 1 -D is preferablyWith L 2 Is thatFor example, said L 1 D may beWith L 2 Is-> For example, said L 1 -D may be->
In a preferred embodiment of the invention, m is preferably 7 to 8, more preferably 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7 or 7.8.
In a preferred embodiment of the present invention, when R is 1 Is substituted or unsubstituted C 1 ～C 10 In the case of alkyl, said C 1 ～C 10 Alkyl is preferably C 1 ～C 4 Alkyl is further preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, most preferably methyl or ethyl.
In a preferred embodiment of the present invention, when R is 1 Is substituted or unsubstituted C 6 ～C 14 Aryl, the C 6 ～C 14 Aryl is preferably phenyl, naphthyl or anthracenyl, more preferably phenyl.
In a preferred embodiment of the present invention, R is 1-1 、R 1-2 And R is 1-3 Independently, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl is preferred, and methyl is further preferred.
In a preferred embodiment of the invention, R 1 In C when substituted 1 ～C 10 The substituent in the alkyl group being hydroxy-substituted C 1 ～C 4 In the case of alkoxy, said C 1 ～C 4 The alkoxy group is preferably methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy or tert-butoxy, more preferably ethoxy.
In a preferred embodiment of the present invention, when R is 3 is-C (=O) R 3-1 When said R is 3-1 Preferably hydroxy-substituted methyl, hydroxy-substituted ethyl,Hydroxy-substituted n-propyl, hydroxy-substituted isopropyl, hydroxy-substituted n-butyl, hydroxy-substituted isobutyl, or hydroxy-substituted tert-butyl, with hydroxy-substituted methyl being further preferred.
In a preferred embodiment of the present invention, said L 3 Preferably is
In a preferred embodiment of the invention, said L is independently preferably a glycine residue or a phenylalanine residue; said p is preferably 4. The (L) p is preferablyWherein the g-terminus is attached to the carbonyl group in formula I.
In a preferred embodiment of the present invention, R is 1 Preferably substituted or unsubstituted C 1 ～C 10 Alkyl, or substituted or unsubstituted C 6 ～C 14 An aryl group; said substituted C 1 ～C 10 Alkyl and substituted C 6 ～C 14 The substituents in the aryl group are preferably one or more groups selected from the group consisting of the following, when multiple substituents are present, the substituents are the same or different: -NR 1- 1 R 1-2 、-S(O) 2 R 1-3 And hydroxy-substituted C 1 ～C 4 An alkoxy group.
In a preferred embodiment of the present invention, R is 2 Preferably hydrogen.
In a preferred embodiment of the invention, said n 1 Preferably 2.
In a preferred embodiment of the invention, said L' is independently preferably a valine residue, an alanine residue, a lysine residue, a phenylalanine residue or a citrulline residue; said p' is preferably 2. Said (L ') p' is preferablyWherein the h-terminus is attached to the carbonyl group in formula II.
In a preferred embodiment of the invention, thePreferably is
In a preferred embodiment of the invention, thePreferably is
In a preferred embodiment of the present invention, said L 2 Preferably is
In a preferred embodiment of the invention, said n 2 Preferably 1.
In a preferred embodiment of the invention, when L 1 -D isWhen said L 2 Preferably is
In a preferred embodiment of the present invention, R is 3 Preferably, -C (=O)R 3-1 。
In a preferred embodiment of the invention, when L 1 -D is In this case, said (L ') p' is preferably +.>
In a preferred embodiment of the invention, when D isWhen L 1 -D is preferably
In a preferred embodiment of the present invention, the antibody conjugated drug wherein the Ab is Trastuzumab, an anti-HER 2 antibody; the D is a cytotoxic drug; m is 2-8;
the L is 1 The structure of (C) is shown as a formula I or II;
the L is independently a phenylalanine residue or a glycine residue; p is 0 to 4;
said R is 1 Is substituted or unsubstituted C 1 ～C 10 Alkyl, or substituted or unsubstituted C 6 ～C 14 An aryl group; the substituted C 1 ～C 10 Alkyl and substituted C 6 ～C 14 The substituents in the aryl group are one or more groups selected from the group consisting of the following, when multiple substituents are present, the substituents are the same or different: -NR 1-1 R 1-2 、-S(O) 2 R 1-3 And hydroxy-substituted C 1 ～C 4 An alkoxy group; said R is 1-1 Said R 1-2 And R is as described 1-3 Independently C 1 ～C 4 An alkyl group;
said R is 2 Is hydrogen;
said n 1 Is 2;
the L' is independently a valine residue, an alanine residue, a lysine residue, a phenylalanine residue or a citrulline residue; the p' is 2-4;
when said L 1 When linked to an amino group in said cytotoxic drug, the hydrogen on said amino group is not substituted or is replaced by an R 3 Substitution; said R is 3 is-C (=O) R 3-1 ；R 3-1 C substituted by hydroxy 1 ～C 4 An alkyl group;
the L is 2 Is thatSaid n 2 1 to 8;
the L is 3 Is that
The amino acid sequence of the light chain in the anti-HER 2 antibody Trastuzumab is preferably shown as SEQ ID No.5 in a sequence table, and the amino acid sequence of the heavy chain is preferably shown as SEQ ID No.6 in the sequence table.
In a preferred embodiment of the present invention, the antibody conjugated drug wherein the Ab is Trastuzumab, an anti-HER 2 antibody; the D isM is 7-8;
the L is 1 The structure of (C) is shown as a formula I or II;
the L is independently a phenylalanine residue or a glycine residue; p is 4;
said R is 1 Is substituted or unsubstituted C 1 ～C 4 Alkyl, or substituted or unsubstituted C 6 ～C 14 An aryl group; the substituted C 1 ～C 4 Alkyl and substituted C 6 ～C 14 The substituents in the aryl group are one or more groups selected from the group consisting of the following, when multiple substituents are present, the substituents are the same or different: -N (CH) 3 ) 2 、-S(O) 2 CH 3 And hydroxy-substituted ethoxy;
said R is 2 Is hydrogen;
said n 1 Is 2;
the L' is independently a valine residue, an alanine residue, a lysine residue, a phenylalanine residue or a citrulline residue; the p' is 2-4;
when said L 1 When linked to an amino group in said cytotoxic drug, the hydrogen on said amino group is not substituted or is replaced by an R 3 Substitution; said R is 3 is-C (=O) R 3-1 ；R 3-1 C substituted by hydroxy 1 ～C 4 An alkyl group;
the L is 2 Is thatSaid n 2 1 to 8;
the L is 3 Is that
And when L 1 -D isWhen said L 2 Is->
The amino acid sequence of the light chain in the anti-HER 2 antibody Trastuzumab is preferably shown as SEQ ID No.5 in a sequence table, and the amino acid sequence of the heavy chain is preferably shown as SEQ ID No.6 in the sequence table.
In a preferred embodiment of the present invention, the antibody conjugated drug wherein the Ab is Trastuzumab, an anti-HER 2 antibody; l (L) 1 -D ism is 7-8;
the L is 1 The structure of (C) is shown as a formula I or II;
the L is independently a phenylalanine residue or a glycine residue; p is 4;
said R is 1 Is substituted or unsubstituted C 1 ～C 4 Alkyl, or substituted or unsubstituted C 6 ～C 14 An aryl group; the substituted C 1 ～C 4 Alkyl or substituted C 6 ～C 14 The substituents in the aryl group are one or more groups selected from the group consisting of the following, when multiple substituents are present, the substituents are the same or different: -N (CH) 3 ) 2 、-S(O) 2 CH 3 And hydroxy-substituted ethoxy;
said R is 2 Is hydrogen;
said n 1 Is 2;
the L' is independently a valine residue, an alanine residue, a lysine residue, a phenylalanine residue or a citrulline residue; the p' is 2-4;
said R is 3 is-C (=O) R 3-1 ；R 3-1 Methyl substituted with hydroxy;
the L is 2 Is thatSaid n 2 1 to 8;
the L is 3 Is that
And, when L 1 -D isIn the case of said formula (L ') p' is +.>When L 1 -D is->When said L 2 Is that
The amino acid sequence of the light chain in the anti-HER 2 antibody Trastuzumab is preferably shown as SEQ ID No.5 in a sequence table, and the amino acid sequence of the heavy chain is preferably shown as SEQ ID No.6 in the sequence table.
In a preferred embodiment of the present invention, the antibody-conjugated drug is preferably a compound shown in any one of the following:
wherein Ab is anti-HER 2 antibody Trastuzumab, anti-B7-H3 antibody P2E5, anti-Claudin 18.2 antibody IMAB362 or anti-Trop 2 antibody RS7, and m is 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7 or 7.8; the amino acid sequence of the light chain in the anti-HER 2 antibody Trastuzumab is preferably shown as SEQ ID No.5 in a sequence table, and the amino acid sequence of the heavy chain is preferably shown as SEQ ID No.6 in the sequence table; the amino acid sequence of the light chain in the anti-B7-H3 antibody P2E5 is preferably shown as SEQ ID No.7 in a sequence table, and the amino acid sequence of the heavy chain is preferably shown as SEQ ID No.8 in the sequence table; the Claudin18.2 antibody IMAB362The amino acid sequence of the light chain is preferably shown as SEQ ID No.1 in the sequence table, and the amino acid sequence of the heavy chain is preferably shown as SEQ ID No.2 in the sequence table; the amino acid sequence of the light chain in the anti-Trop 2 antibody RS7 is preferably shown as SEQ ID No.3 in the sequence table, and the amino acid sequence of the heavy chain is preferably shown as SEQ ID No.4 in the sequence table.
In a preferred embodiment of the present invention, the antibody-conjugated drug is preferably a compound shown in any one of the following:
further preferably +.>
Even more preferably +.> Wherein Ab is the anti-HER 2 antibody Trastuzumab; the amino acid sequence of the light chain in the anti-HER 2 antibody Trastuzumab is shown as SEQ ID No.5 in a sequence table, and the amino acid sequence of the heavy chain is shown as SEQ ID No.6 in the sequence table.
The invention also provides a linker-drug conjugate, which has a general structural formula of L 4 -L 2 -L 1 -D, wherein L 4 Is thatL 2 、L 1 And D is as defined above, L 2 And with L 4 Is connected to the d terminal of (c).
In a preferred embodiment of the invention, the linker-drug conjugate is preferably any one of the compounds shown below:
the invention provides a preparation method of the antibody coupling drug, which comprises the following steps of coupling the linker-drug conjugate with an antibody.
In the present invention, the conditions and operations of the coupling may be those conventional in the art.
The invention also provides a pharmaceutical composition comprising the antibody-conjugated drug and a pharmaceutically acceptable carrier.
The invention also provides an application of the antibody coupling medicament or the pharmaceutical composition in preparing medicaments for preventing or treating cancers. The cancer is preferably stomach cancer, breast cancer, non-small cell lung cancer, urothelial cancer or pancreatic cancer.
The invention also provides a pharmaceutical preparation comprising the antibody-conjugated drug.
In the present invention, m represents the molar ratio of cytotoxic drug molecule to Ab (also known as DAR, i.e. drug antibody coupling ratio), m may be an integer or a fraction, preferably understood as: the average molar ratio of the drug molecules in the antibody-conjugated drug obtained after the conjugation of the single monoclonal antibody molecule and the cytotoxic drug to the monoclonal antibody molecules can be generally determined by means of Hydrophobic chromatography (Hydrophobic-Interaction Chromatography, HIC), polyacrylamide-SDS gel electrophoresis (SDS-PAGE), liquid phase mass spectrometry (liquid chromatograph-mass spectrometer, LC-MS) and the like.
Antibodies of the invention may be prepared using techniques well known in the art, such as hybridoma methods, recombinant DNA techniques, phage display techniques, synthetic techniques, or combinations thereof, or other techniques known in the art.
Variants refer to mutants of the amino acid sequence of an antibody, as well as covalent derivatives of the native polypeptide, provided that the biological activity equivalent to the native polypeptide is retained. Amino acid sequence mutants generally differ from the native amino acid sequence in that one or more amino acids in the native amino acid sequence are replaced or one or more amino acids are deleted and/or inserted in the polypeptide sequence. Deletion mutants include fragments of the native polypeptide and N-and/or C-terminal truncation mutants. Typically, amino acid sequence mutants have at least 70%, 75%, 80%, 85%, 90%, 95%, 98% or more than 99% homology to the native sequence.
The term "treatment" or its equivalent, when used in reference to, for example, cancer, refers to a procedure or process used to reduce or eliminate the number of cancer cells in a patient or to alleviate symptoms of cancer. "treating" of cancer or another proliferative disorder does not necessarily mean that the cancer cells or other disorder will actually be eliminated, that the number of cells or disorders will actually be reduced or that the symptoms of the cancer or other disorder will actually be reduced. In general, methods of treating cancer are performed even with low likelihood of success, but are still considered to induce an overall beneficial course of action, given the patient's medical history and estimated survival expectancy.
The term "pharmaceutically acceptable carrier" refers to any formulation or carrier medium representative of a carrier capable of delivering an effective amount of the active agents of the present invention, which does not interfere with the biological activity of the active agents and which does not have toxic or side effects to the host or patient, including water, oils, vegetables and minerals, cream bases, lotion bases, ointment bases, and the like. Such matrices include suspending agents, viscosity enhancers, transdermal enhancers, and the like. Their formulations are well known to those skilled in the cosmetic or topical pharmaceutical arts.
The above preferred conditions can be arbitrarily combined on the basis of not deviating from the common knowledge in the art, and thus, each preferred embodiment of the present invention can be obtained.
The reagents and materials used in the present invention are commercially available.
The invention has the positive progress effects that: the novel antibody coupling medicine can realize the wide application of cytotoxic medicines in the field of ADC and treat tumor patients with microtubule ADC drug resistance.
Detailed Description
The invention is further illustrated by means of the following examples, which are not intended to limit the scope of the invention. The experimental methods, in which specific conditions are not noted in the following examples, were selected according to conventional methods and conditions, or according to the commercial specifications.
Preparation example 1 preparation of DS-8201a linker-drug conjugate GGFG-Dxd
The compound GGFG-Dxd (structure below) was synthesized by known methods reported in reference to WO2015146132A1, ESI-MS m/z:1034.5 (M+H), 1 H-NMR(400MHz,DMSO-d 6 )δ8.61(t,J＝6.4Hz,1H),8.50(d,J＝8.5Hz,1H),8.28(t,J＝5.1Hz,1H),8.11(d,J＝7.5Hz,1H),8.05(t,J＝5.7Hz,1H),7.99(t,J＝5.9Hz,1H),7.77(d,J＝11.0Hz,1H),7.31(s,1H),7.25–7.16(m,5H),6.98(s,2H),6.51(s,1H),5.59(dt,J＝7.4,4.1Hz,1H),5.41(s,2H),5.20(s,2H),4.64(d,J＝6.1Hz,2H),4.53–4.40(m,1H),4.02(s,2H),3.74–3.37(m,8H),3.18–3.00(m,2H),3.04–2.97(m,1H),2.77(dd,J＝13.5,9.4Hz,1H),2.38(s,3H),2.19(dd,J＝14.9,8.5Hz,2H),2.11–2.05(m,2H),1.86(dd,J＝14.0,6.7Hz,2H),1.45(s,4H),1.20–1.14(m,2H),0.87(t,J＝7.1Hz,3H).
EXAMPLE 1 Synthesis of Compound DX01
Step 1 synthesis of compound IIIC:
commercial compound IIIA (5.58 g,10 mmol) and commercial IIIB (3.16 g,10 mmol) were mixed in 100mL anhydrous N, N-dimethylformamide, HATU (1.14 g,3.0 mmol) was added, and triethylamine 10mL reacted at room temperature for 2h. After the reaction, the solvent is removed by reduced pressure distillation, and the crude product is subjected to silica gel column chromatography [ chloroform: methanol=10:1 (v/v) ] to afford the title compound IIIC (7.71 g, yield 90%), ESI-MS m/z:857.1 (M+H).
Step 2 synthesis of compound IIID:
compound IIIC (7.71 g,9 mmol) was dissolved in dichloromethane and 9mL of trifluoroacetic acid was added and reacted at room temperature for 0.5h. After the reaction is finished, the solvent is removed by reduced pressure distillation, and the crude product is subjected to silica gel column chromatography [ dichloromethane: methanol=10:1 (v/v) ] to afford the trifluoroacetate salt of the title compound IIID (5.90 g, yield 90%), ESI-MS m/z:614.9 (M+H).
Step 3 synthesis of compound IIIE:
tert-butyl 2-glycolate (2.5 g,18.9 mmol) and di (p-nitrophenyl) carbonate (6.3 g,20.8 mmol) were mixed and dissolved in 200mL anhydrous DMF, 25mL triethylamine was added and reacted at room temperature for 2 hours. After completion of the reaction of the starting materials by liquid chromatography, trifluoroacetic acid salt (7 g,9.6 mmol) of IIID was added thereto, and the reaction was continued for 1 hour. After the reaction, the solvent was removed by distillation under reduced pressure, 150mL of water was added, extraction was performed three times with ethyl acetate (100 mL each time), the organic phases were combined after separation, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated, and the crude product was purified by silica gel column chromatography [ dichloromethane: ethyl acetate=10:1 (v/v) ] to afford the title compound IIIE (6.8 g, 92% yield), ESI-MS m/z:773.5 (M+H).
Step 4 synthesis of compound IIIF:
compound IIIE (7.73 g,10 mmol) was dissolved in 150mL dichloromethane and 9mL trifluoroacetic acid was added and reacted at room temperature for 0.5h. After the reaction is finished, the solvent is removed by reduced pressure distillation, and the crude product is subjected to silica gel column chromatography [ dichloromethane: methanol=10:1 (v/v) ] to afford the title compound IIIF (5.73 g, 80% yield), ESI-MS m/z:717.2 (M+H).
Step 5 synthesis of compound IIIG:
compound IIIF (5.73 g,8 mmol) was mixed with commercially available Exatecan mesylate (4.5 g,8 mmol) in 30mL anhydrous DMF and HATU (3.8 g,10 mmol), triethylamine 2mL and reacted at room temperature for 2h. After the reaction, the solvent is removed by reduced pressure distillation, and the crude product is subjected to silica gel column chromatography [ chloroform: methanol=10:1 (v/v) ] to afford the title compound IIIG (7.89 g, yield 87%), ESI-MS m/z:1134.1 (M+H).
Step 6 synthesis of compound DX 01:
compound IIIG (1 g,0.929 mmol) was dissolved in 20mL anhydrous DMF and 0.5mL 1, 8-diazabicyclo undec-7-ene was added and reacted at room temperature for 1 hour. After the reaction of the starting materials was completed, succinimidyl 6- (maleimide) hexanoate (428.5 mg,1.39 mmol) was directly added thereto and stirred at room temperature for 1 hour. The solvent was distilled off under reduced pressure, and the crude product was purified by silica gel column chromatography [ chloroform: methanol=8:1 (v/v) ] to afford the title compound (0.96 g, 73% yield), ESI-MS m/z:1105.3 (M+H).
Example 2
Synthesis of DX02, DX03, DX04
Referring to example 1, compound IIIA was reacted with a different commercially available amino fragment at step 1, and the subsequent procedure was the same as in example 1, to finally obtain compound DX02-DX04.
Compound DX02: pale yellow powder, ESI-MS m/z:1164.3 (m+h); compound DX03: pale yellow powder, ESI-MS m/z:1199.5 (m+h); compound DX04: pale yellow powder, ESI-MS m/z:1169.2(M+H)。
EXAMPLE 3 Synthesis of Compounds DX05, DX06
Step 1 Synthesis of Compound DXD-1
Commercial compound DXD (1 g,2.03 mmol) and t-butyldimethylchlorosilane (0.46 g,3.05 mmol) were dissolved in 20mL of anhydrous dichloromethane, 1mL of pyridine was added, and the mixture was stirred at room temperature for 2 hours. After the reaction, the solvent was evaporated under reduced pressure, and the crude product was purified by column chromatography [ dichloromethane: methanol=50:1 (v/v) ] to afford the title compound (1.11 g, yield 90%), ESI-MS m/z:608.1 (M+H).
Step 2 Synthesis of intermediate IX
1 equivalent of compound DXD-1 was dissolved in anhydrous methylene chloride, 0.5 equivalent of triphosgene was added, and then 2 equivalents of p-dimethylaminopyridine was added to react at room temperature for 1 hour. Intermediate V (R represents different substituents corresponding to DX05 and DX06 respectively) was added and then reacted for 0.5 hour at room temperature, and 10% trifluoroacetic acid was added for 1 hour to remove the silicon protection. Then evaporating the solvent under reduced pressure, purifying the crude product by column chromatography to obtain an intermediate IX, and obtaining a target product (wherein R in DX05 is methyl; R in DX06 is R) by a subsequent reaction step according to example 1). Compound DX05: pale yellow solid, ESI-MS m/z:1107.3 (m+h); compound DX06: colorless oil, ESI-MS m/z:1181.2 (M+H).
EXAMPLE 4 Synthesis of Compounds DX07 and DX08
Intermediate VIII-5 and IX-5 were obtained as described in either preparation example 1 or example 2, and then treated with 1, 8-diazabicyclo undec-7-ene as described in step 6 of example 1 to react with commercially available starting material B to give the desired product. Compound DX07: colorless oil, ESI-MS m/z:1305.5 (m+h); compound DX08: pale yellow oil, ESI-MS m/z:1305.3 (M+H).
EXAMPLE 5 Synthesis of Compounds DX09, DX10
Step 1 synthesis of compound 2:
commercial Compound 1 (10 g,26.4 mmol) was mixed with 4-azidobutyric acid (5.11 g,39.6 mmol) and dissolved in 100mL anhydrous DMF, EEDQ (13.1 g,52.8 mmol) was added and reacted for 5 hours at room temperature. After the reaction, the solvent was distilled off under reduced pressure, and then subjected to silica gel column chromatography [ dichloromethane: methanol=10:1 (v/v) ] to afford the title compound (11.8 g, 91% yield), ESI-MS m/z:491.3 (M+H).
Step 2 synthesis of compound 3:
compound 2 (10 g,20.4 mmol) was dissolved in 100mL anhydrous DMF, cooled to 0℃and thionyl chloride (1.8 mL,24.5 mmol) was slowly added and the reaction continued at this temperature for 30 min. After the reaction, ice water was slowly added to quench the reaction, and a solid was precipitated, filtered, and the cake was washed with water, methyl t-butyl ether, and dried under reduced pressure to give the title compound (8.6 g, yield 83%) ESI-MS m/z:509.1 (M+H).
Step 3 synthesis of compound 4:
compound 3 (1 g,1.97 mmol) was mixed with commercially available Exatecan mesylate (1.12 g,1.97 mmol) and dissolved in 50mL anhydrous DMF and triethylamine (1 mL) was added and reacted at room temperature for 2 hours. After the reaction is finished, the solvent is removed by reduced pressure distillation, and the crude product is subjected to silica gel column chromatography [ dichloromethane: methanol=10:1 (v/v) ] to afford the title compound (1.1 g, 63% yield), ESI-MS m/z:908.1 (M+H).
Step 4 synthesis of compound 5:
compound 4 (1 g,1.10 mmol) was mixed with glycolic acid (83.7 mg,1.10 mmol) and dissolved in 20mL anhydrous DMF, HATU (836 mg,2.20 mmol) was added, triethylamine 1mL and reacted at room temperature for 1.5 h. After the reaction, the solvent was distilled off under reduced pressure, and the mixture was purified by silica gel column chromatography [ dichloromethane: methanol=10:1 (v/v) ] to afford the title compound (542 mg, yield 51%), ESI-MS m/z:966.4 (M+H).
Step 5 synthesis of compound DX 09:
compound 5 (500 mg,0.518 mmol) and commercially available starting material C (128.6 mg,0.518 mmol) were dissolved in 10mL anhydrous DMF and a catalytic amount of cuprous bromide was added to react for 1 hour at room temperature. After the reaction, the solvent was distilled off under reduced pressure, and the mixture was purified by silica gel column chromatography [ chloroform: methanol=10:1 (v/v) ] to afford the title compound (546 mg, yield 87%), ESI-MS m/z:1214 (M+H).
Synthesis of Compound DX10 referring to the synthesis of DX09, the raw material compound 5 was changed to Compound 4 to give Compound DX10 as pale yellow solid, ESI-MS m/z:1116.3 (M+H).
EXAMPLE 6 Synthesis of Compounds DX11, DX12
Referring to example 5, the dipeptide starting material was replaced with the corresponding dipeptide starting material commercially available in the initial step, and the subsequent steps were the same as in example 5. Compound DX11: pale yellow solid, ESI-MS m/z:1088.1 (M+H). Compound DX12: pale yellow foamy solid, ESI-MS m/z:1193.3 (M+H).
Example 7 general procedure for ADC preparation
anti-HER 2 antibody Trastuzumab (15 mg/ml concentration) was replaced with G25 desalting column to 50mM PB/1.0mM EDTA buffer (pH 7.0), 15 equivalents TECP was added, stirred at 37 ℃ for 2 hours to allow complete opening of the inter-chain disulfide bonds, then the reduced antibody solution pH was adjusted to 6.0 using phosphoric acid, and the water bath temperature was reduced to 25 ℃ for the coupling reaction. The linker-drug conjugates prepared in preparation example 1 and examples 1 to 6 were dissolved in DMSO, respectively, from which 12 equivalents of linker-drug conjugate linker were pipetted dropwise into the reduced antibody solution, and DMSO was added to a final concentration of 10% (v/v), and the reaction was stirred at 25 ℃ for 0.5 hour, after which the sample was filtered using a 0.22um membrane. Uncoupled small molecules were removed by purification using a tangential flow ultrafiltration system with buffer 50mM PB/1.0mM EDTA solution (pH 6.0), and after purification 6% sucrose was added and stored in a-20deg.C freezer. The absorbance values were measured at 280nm and 370nm, respectively, using the UV method, and DAR values were calculated, with the results shown in Table 1 below. The amino acid sequence of the light chain in the anti-HER 2 antibody Trastuzumab is shown as SEQ ID No.5 in the sequence table, and the amino acid sequence of the heavy chain is shown as SEQ ID No.6 in the sequence table.
TABLE 1 DAR values measured by different antibody conjugated drug (ADC) UV methods
| ADC numbering | Antibodies to | Medicine connector | DAR value |
| DS-8201a | Trastuzumab | GGFG-Dxd | 7.6 |
| ADC-1 | Trastuzumab | DX01 | 7.5 |
| ADC-2 | Trastuzumab | DX02 | 7.4 |
| ADC-3 | Trastuzumab | DX03 | 7.7 |
| ADC-4 | Trastuzumab | DX04 | 7.2 |
| ADC-5 | Trastuzumab | DX05 | 7.1 |
| ADC-6 | Trastuzumab | DX06 | 7.3 |
| ADC-7 | Trastuzumab | DX07 | 7.6 |
| ADC-8 | Trastuzumab | DX08 | 7.5 |
| ADC-9 | Trastuzumab | DX09 | 7.8 |
| ADC-10 | Trastuzumab | DX10 | 7.5 |
| ADC-11 | Trastuzumab | DX11 | 7.5 |
| ADC-12 | Trastuzumab | DX12 | 7.4 |
Effect example 1: in vitro cytotoxic Activity assay
And selecting NCI-N87 cells and SK-BR-3 cells which are stably transfected and highly express Her2 as cell strains for detecting the in vitro activity of the experiment, and observing the quantitative effect of different antibody coupling drugs on cell killing. Seed density of each cell was initially selected: 2X 10 3 cell/hole, and carrying out cytotoxic activity measurement after 16-24 hours; then, the final concentration of the antibody-coupled drug prepared in test example 7 after sample addition was set to 5000nM as the initial concentration, 10 concentrations (4-10-fold dilution) of the designed series of 5000-0.006 nM were observed for 96 hours of killing (or inhibition) change,luminescent Cell Viability Assay chemiluminescence staining, and IC50 was calculated after reading fluorescence data. From the results of the activity test, all ADCs showed some antitumor activity, part of the ADCs exceeded DS-8201a, part of the ADCs were equivalent to DS-8201a, and part of the ADCs were significantly weaker than DS-8201a, as shown in Table 2.
TABLE 2 in vitro cytotoxic Activity of different ADCs
Effect example 2: stability test
This example evaluates the stability of the antibody-conjugated drug of example 7 in a weakly acidic environment. Specifically, in this example, a part of the antibody-conjugated drug of example 7 was added to a buffer solution of pH 4.5, placed in a water bath at 37 ℃ for 1, 2, 3, 4 days with an internal standard (irinotecan as an internal standard substance) and then the release amount of the free drug was detected by high performance liquid chromatography, and the results are shown in table 3.
Table 3 evaluation of stability of partial ADC in weakly acidic environment
The stability results show that the ADC obtained by adopting the novel technical scheme has stronger stability than DS-8201a in a weak acid environment, and various normal tissues in a human body are weak acid, so that the ADC is expected to show better safety and similar effectiveness than DS-8201 a.
Claims (15)
1. An antibody conjugated drug has a structural general formula of Ab- (L) 3 -L 2 -L 1 -D) m ；
Wherein Ab is an antibody;
d is a cytotoxic drug;
m is 2-8;
L 1 the structure of (C) is shown as a formula II; wherein the a-terminus is linked to said cytotoxic agent, the e-terminus is linked to said L 2 Is connected with the c end of the (C) chain,
l' is independently a phenylalanine residue, glycine residue, glutamic acid residue, aspartic acid residue, cysteine residue, glutamic acid residue, histidine residue, isoleucine residue, leucine residue, lysine residue, methionine residue, proline residue, serine residue, threonine residue, tryptophan residue, tyrosine residue or valine residue; p' is 2-4;
L 2 is that Wherein, the c end and L 1 Is connected to the e terminal, the f terminal and the L terminal 3 Is connected with d end of n 2 Independently 1 to 8;
L 3 is thatWherein the b-terminus is linked to said Ab, the d-terminus is linked to said L 2 Is connected to the f terminal of (c).
2. The antibody-conjugated drug of claim 1, wherein,
the antibody is anti-HER 2 antibody Trastuzumab or a variant thereof, anti-B7-H3 antibody P2E5 or a variant thereof, anti-Claudin 18.2 antibody IMAB362 or a variant thereof, or anti-Trop 2 antibody RS7 or a variant thereof, preferably anti-HER 2 antibody Trastuzumab or a variant thereof, and more preferably anti-HER 2 antibody Trastuzumab; the amino acid sequence of the light chain in the anti-HER 2 antibody Trastuzumab is preferably shown as SEQ ID No.5 in a sequence table, and the amino acid sequence of the heavy chain is preferably shown as SEQ ID No.6 in the sequence table; the amino acid sequence of the light chain in the anti-B7-H3 antibody P2E5 is preferably shown as SEQ ID No.7 in a sequence table, and the amino acid sequence of the heavy chain is preferably shown as SEQ ID No.8 in the sequence table; the amino acid sequence of the light chain in the Claudin18.2 antibody IMAB362 is preferably shown as SEQ ID No.1 in a sequence table, and the amino acid sequence of the heavy chain is preferably shown as SEQ ID No.2 in the sequence table; the amino acid sequence of the light chain in the anti-Trop 2 antibody RS7 is preferably shown as SEQ ID No.3 in a sequence table, and the amino acid sequence of the heavy chain is preferably shown as SEQ ID No.4 in the sequence table;
and/or, said L 3 The thiol on the antibody is connected with the segment b of the antibody in the form of thioether bond;
and/or D is a hydroxyl-containing or amino-containing cytotoxic drug, preferably a hydroxyl-containing or amino-containing topoisomerase inhibitor, more preferably a hydroxyl-containing or amino-containing topoisomerase I inhibitor, even more preferably camptothecin or a derivative thereof, most preferablyWhen said L 1 When linked to an amino group in said cytotoxic drug, the hydrogen on said amino group is not substituted or is replaced by an R 3 Substitution; said R is 3 Is C 1 ～C 6 Alkyl, C 3 ～C 8 Cycloalkyl, C 6 ～C 14 Aryl, 5-to 10-membered heteroaryl or-C (=o) R 3-1 The method comprises the steps of carrying out a first treatment on the surface of the The hetero atom in the 5-10 membered heteroaryl is selected from one or more of N, O and S, and the number of the hetero atom is 1, 2, 3 or 4; r is R 3-1 C substituted by hydroxy 1 ～C 4 An alkyl group;
and/or m is 7 to 8, preferably 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7 or 7.8;
and/or, said L 3 Is that
And/or, the L' is independently valine residue, alanine residue, lysine residue, phenylalanine residue or citrulline residue; said p' is preferably 2; said (L ') p' is preferably Wherein the h-terminus is attached to a carbonyl group in formula II；
And/or, said L 2 Is that
And/or, the n 2 1.
3. The antibody-conjugated drug of claim 2, wherein,
said R is 3 is-C (=O) R 3-1 The method comprises the steps of carrying out a first treatment on the surface of the Said R is 3-1 Preferably a hydroxy-substituted methyl group, a hydroxy-substituted ethyl group, a hydroxy-substituted n-propyl group, a hydroxy-substituted isopropyl group, a hydroxy-substituted n-butyl group, a hydroxy-substituted isobutyl group, or a hydroxy-substituted tert-butyl group, more preferably a hydroxy-substituted methyl group;
and/or when L 1 -D isWhen said L 2 Is->
And/or when L 1 -D isIn the case of said formula (L ') p' is +.>
4. The antibody-conjugated drug of claim 1, wherein,
the Ab is an anti-HER 2 antibody Trastuzumab; the D is a cytotoxic drug; m is 2-8;
the L is 1 The structure of (C) is shown as a formula II;
the L' is independently a valine residue, an alanine residue, a lysine residue, a phenylalanine residue or a citrulline residue; the p' is 2-4;
when said L 1 When linked to an amino group in said cytotoxic drug, the hydrogen on said amino group is not substituted or is replaced by an R 3 Substitution; said R is 3 is-C (=O) R 3-1 ；R 3-1 C substituted by hydroxy 1 ～C 4 An alkyl group;
the L is 2 Is thatSaid n 2 1 to 8;
the L is 3 Is that
The amino acid sequence of the light chain in the anti-HER 2 antibody Trastuzumab is preferably shown as SEQ ID No.5 in a sequence table, and the amino acid sequence of the heavy chain is preferably shown as SEQ ID No.6 in the sequence table.
5. The antibody-conjugated drug of claim 4, wherein,
the Ab is an anti-HER 2 antibody Trastuzumab; the D isM is 7-8;
the L is 1 The structure of (C) is shown as a formula II;
the L' is independently a valine residue, an alanine residue, a lysine residue, a phenylalanine residue or a citrulline residue; the p' is 2-4;
when said L 1 When linked to an amino group in said cytotoxic drug, the hydrogen on said amino group is not substituted or is replaced by an R 3 Substitution; said R is 3 is-C (=O) R 3-1 ；R 3-1 C substituted by hydroxy 1 ～C 4 An alkyl group;
the L is 2 Is thatSaid n 2 1 to 8;
the L is 3 Is that
And when L 1 -D isWhen said L 2 Is->
6. The antibody-conjugated drug of claim 5,
the Ab is an anti-HER 2 antibody Trastuzumab; l (L) 1 -D is M is 7-8;
the L is 1 The structure of (C) is shown as a formula II;
the L' is independently a valine residue, an alanine residue, a lysine residue, a phenylalanine residue or a citrulline residue; the p' is 2-4;
said R is 3 is-C (=O) R 3-1 ；R 3-1 Methyl substituted with hydroxy;
the L is 2 Is thatSaid n 2 1 to 8;
the L is 3 Is that
And when L 1 -D isIn the case of (L ') p' is
7. The antibody-conjugated drug of claim 1, wherein the antibody-conjugated drug is a compound as shown in any one of the following:
the Ab in the kit is anti-HER 2 antibody Trastuzumab, anti-B7-H3 antibody P2E5, anti-Claudin 18.2 antibody IMAB362 or anti-Trop 2 antibody RS7, and m is 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7 or 7.8, the amino acid sequence of a light chain in the anti-HER 2 antibody Trastuzumab is preferably shown as SEQ ID No.5 in a sequence table, and the amino acid sequence of a heavy chain is preferably shown as SEQ ID No.6 in the sequence table; the amino acid sequence of the light chain in the anti-B7-H3 antibody P2E5 is preferably shown as SEQ ID No.7 in a sequence table, and the amino acid sequence of the heavy chain is preferably shown as SEQ ID No.8 in the sequence table; the amino acid sequence of the light chain in the Claudin18.2 antibody IMAB362 is preferably shown as SEQ ID No.1 in a sequence table, and the amino acid sequence of the heavy chain is preferably shown as SEQ ID No.2 in the sequence table; the amino acid sequence of the light chain in the anti-Trop 2 antibody RS7 is preferably shown as SEQ ID No.3 in the sequence table, and the amino acid sequence of the heavy chain is preferably shown as SEQ ID No.4 in the sequence table.
8. The antibody-conjugated drug of claim 7, wherein the antibody-conjugated drug is a compound as shown in any one of the following:
further preferably +.> Even more preferably +.> Wherein Ab is the anti-HER 2 antibody Trastuzumab.
9. The antibody-conjugated drug of claim 8, wherein the antibody-conjugated drug is a compound as shown in any one of the following:
further preferably +.> Even more preferably +.> Wherein Ab is the anti-HER 2 antibody Trastuzumab; the amino acid sequence of the light chain in the anti-HER 2 antibody Trastuzumab is shown as SEQ ID No.5 in the sequence table, and the amino acid sequence of the heavy chain is shown as SEQ ID No.6 in the sequence table.
10. A linker-drug conjugate has the general structural formula L 4 -L 2 -L 1 -D, wherein L 4 Is that L 2 、L 1 And D is as defined in any one of claims 1 to 7, L 2 And with L 4 Is connected to the d terminal of (c).
11. The linker-drug conjugate of claim 10 which is any one of the compounds shown below:
12. a method of preparing an antibody-conjugated drug according to any one of claims 1 to 9, comprising the step of conjugating said antibody to a linker-drug conjugate according to claim 10 or 11.
13. A pharmaceutical composition comprising the antibody-conjugated drug of any one of claims 1-9 and a pharmaceutically acceptable carrier.
14. Use of an antibody-conjugated drug according to any one of claims 1 to 9, or a pharmaceutical composition according to claim 13, in the manufacture of a medicament for the prevention or treatment of cancer.
15. A pharmaceutical formulation comprising the antibody-conjugated drug of any one of claims 1-9.
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| WO2022205111A1 (en) * | 2021-03-31 | 2022-10-06 | 上海复旦张江生物医药股份有限公司 | Method for preparing exatecan derivative, and intermediate thereof |
| WO2023016564A1 (en) * | 2021-08-12 | 2023-02-16 | 上海才致药成生物科技有限公司 | GPC3-TARGETED ANTIBODY INTERFERON α FUSION PROTEIN AND USE THEREOF |
| WO2023061457A1 (en) * | 2021-10-14 | 2023-04-20 | 百奥泰生物制药股份有限公司 | Antibody-drug conjugate and application thereof |
| WO2023232145A1 (en) * | 2022-06-02 | 2023-12-07 | 华东师范大学 | Small molecule of homocamptothecins and use thereof |
| WO2024008102A1 (en) * | 2022-07-05 | 2024-01-11 | Wuxi Xdc (Shanghai) Co., Ltd. | Linker for conjugation |
| WO2024026323A1 (en) * | 2022-07-26 | 2024-02-01 | Zeno Management, Inc. | Immunoconjugates and methods |
| WO2024125627A1 (en) * | 2022-12-16 | 2024-06-20 | 上海齐鲁制药研究中心有限公司 | Camptothecin compound, preparation method therefor, and use thereof |
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| PL3903829T3 (en) * | 2009-02-13 | 2023-08-14 | Immunomedics, Inc. | Immunoconjugates with an intracellularly-cleavable linkage |
| CN115960111A (en) * | 2012-10-11 | 2023-04-14 | 第一三共株式会社 | Antibody-drug conjugates |
| JP6612738B2 (en) * | 2014-04-10 | 2019-11-27 | 第一三共株式会社 | Anti-HER2 antibody-drug conjugate |
| CN109106951A (en) * | 2017-08-18 | 2019-01-01 | 四川百利药业有限责任公司 | A kind of camptothecine-antibody coupling matter |
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