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CN112138171A - Antibody coupling drug, intermediate thereof, preparation method and application - Google Patents

Antibody coupling drug, intermediate thereof, preparation method and application Download PDF

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CN112138171A
CN112138171A CN201910576227.3A CN201910576227A CN112138171A CN 112138171 A CN112138171 A CN 112138171A CN 201910576227 A CN201910576227 A CN 201910576227A CN 112138171 A CN112138171 A CN 112138171A
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substituted
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antibody
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CN112138171B (en
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鲍彬
郭青松
高贝
张一帆
邱雪飞
杨彤
沈毅珺
张文伯
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SHANGHAI FUDAN-ZHANGJIANG BIO-PHARMACEUTICAL CO LTD
Taizhou Fudan Zhangjiang Pharmaceutical Co Ltd
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SHANGHAI FUDAN-ZHANGJIANG BIO-PHARMACEUTICAL CO LTD
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Priority to CN202311489217.9A priority patent/CN117731798A/en
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    • A—HUMAN NECESSITIES
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    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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    • A61K47/50—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
    • 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
    • A61K47/6835—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
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K47/50—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
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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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    • A61K47/68—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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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Abstract

The invention discloses an antibody conjugate drug, an intermediate thereof, a preparation method and application. The invention discloses an antibody coupling drug with a structural general formula of Ab- (L)3‑L2‑L1‑D)m. The antibody conjugate drug can realize the wide application of cytotoxic drugs, particularly camptothecin in the field of ADC, and treat tumor patients with drug resistance to microtubule ADC.

Description

Antibody coupling drug, intermediate thereof, preparation method and application
Technical Field
The invention belongs to the field of biotechnology and medicine, and particularly relates to an antibody conjugate drug, an intermediate, a preparation method and application thereof.
Background
Antibody-conjugated drugs (ADCs) are one of the hot spots of interest to the pharmaceutical industry in recent years. Because of the unsatisfactory clinical efficacy of many antibody drugs, many industries are increasingly turning their eyes to ADC drugs. Four ADC drugs are currently marketed abroad. Gemtuzumab Ozogamicin (trade name Mylotarg) approved by FDA at 17 days 5.2000 for fevery, was marketed for the treatment of Acute Myelogenous Leukemia (AML) patients with first relapse, above 60 years old, CD33+, who were not suitable for cytotoxic chemotherapy, although this drug was withdrawn from the market in 2010 but re-marketed in 2017, and Inotuzumab Ozogamicin (trade name bestonsa) of homophony was also approved by FDA for the treatment of adult relapse refractory B-cell ALL. Brentuximab Vedotin (trade name Adcetris) developed by the FDA approved Seattle Genetics company is marketed at 19.8.2011 for the treatment of CD30 positive Hodgkin Lymphoma (HL) and the rare disease Systemic Anaplastic Large Cell Lymphoma (SALCL). On day 22/2/2013, ado-trastuzumab emtansine (T-DM1, trade name Kadcyla) developed by Genentech corporation was approved by FDA for marketing, and was mainly used for treating Her2 positive advanced (metastatic) breast cancer. In addition, more than 100 ADC drugs are still in clinical as well as preclinical development stages internationally.
The linker of ADC is an important component, and the stability of the linker not only influences the release of small molecules in tumor cells, but also is related to the toxic and side effects of ADC. Linkers that are too stable are generally not easily released, such as T-DM 1; an excessively labile linker tends to release the drug in the plasma, thereby causing toxicity, such as IMMU-132. The patent (patent number ZL201380053256) disclosed by the first three co-companies invents a very stable linking group in blood plasma, namely, a camptothecin compound is linked with an antibody through a section of enzyme-cleavable tetrapeptide and a self-cleavable aminomethyl ether structure, so that the camptothecin compound has a good anti-tumor effect. Although the structure can keep better stability in plasma, the amino methyl ether structure is unstable in acid, and a plurality of acid environments exist in vivo, including part of tumor tissues and some normal tissues. Thus, ADC releases part of the drug molecule without being endocytosed by the tumor cell, 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 amino methyl ether structure with a carbamide structure to enhance the stability. The other is that the self-cleavage fragment is directly connected to the drug molecule by alkyl, the structure has good stability, and can be rapidly eliminated by 1, 6-after enzyme digestion, thereby releasing the drug. Therefore, the invention can solve the defects of the ADC, simultaneously can keep better biological activity, and obtains 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 the antibody coupling drug, an intermediate, a preparation method and application thereof. The antibody coupling drug can realize the wide application of cytotoxic drugs in the ADC field and treat microtube ADC drug-resistant tumor patients.
The invention solves the technical problems through the following technical scheme.
The invention provides an antibody coupling drug, the structural general formula of which is Ab- (L)3-L2-L1-D)m；
Wherein Ab is an antibody;
d is a cytotoxic drug;
m is 2-8;
L1the structure of (A) is shown as formula I or II; wherein the a-terminus is linked to said cytotoxic agent and the e-terminus is linked to said L2The end c of the first and second terminals are connected,
Figure BDA0002112110080000021
l is independently a phenylalanine residue, a glycine residue, a glutamic acid residue, an aspartic acid residue, a cysteine residue, a glutamic acid residue, a histidine residue, an isoleucine residue, a leucine residue, a lysine residue, a methionine residue, a proline residue, a serine residue, a threonine residue, a tryptophan residue, a tyrosine residue, or a valine residue; p is 0 to 4;
R1is hydrogen, substituted or unsubstituted C1～C10Alkyl, substituted or unsubstituted C6～C14An aryl group, a substituted or unsubstituted 5-to 10-membered heterocyclic group, or a substituted or unsubstituted phenoxy group; said substituted C1～C10Alkyl, substituted C6～C14The 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 the following, and when a plurality of substituents are present, the substituents may be the same or different: halogen, hydroxy, -NR1-1R1-2、-S(O)2R1-3、C1～C4Alkyl radical, C1～C4Alkoxy, hydroxy-substituted C1～C4Alkoxy radical, C3～C8Cycloalkyl, 5-to 8-membered heterocyclic group, C6～C10Aryl 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; r1-1、R1-2And R1-3Independently is C1～C4An alkyl group;
R2independently hydrogen, substituted or unsubstituted C1～C10Alkyl, substituted or unsubstituted C3～C10Cycloalkyl, substituted or unsubstituted C6～C14Aryl, substituted or unsubstituted 5-to 10-membered heteroaryl; said substituted C1～C10Alkyl, substituted C3～C10Cycloalkyl, substituted C6～C14The substituents in the aryl group and the substituted 5-to 10-membered heteroaryl group are one or more groups selected from the group consisting of the following groups, and when a plurality of substituents are present, the substituents are the same or different: halogen, hydroxy, C1～C4Alkyl radical, C1～C4Alkoxy radical, C3～C8Cycloalkyl, 5-to 8-membered heterocyclic group, C6～C10Aryl and 5-to 10-membered heteroaryl; the heteroatoms in the 5-to 8-membered heterocyclic group, the 5-to 10-membered heteroaryl group and the substituted or unsubstituted 5-to 10-membered heterocyclic group are selected from one or more of N, O and S, and the number of the heteroatoms is 1, 2, 3 or 4;
n1is 2, 3 or 4;
l' is independently a phenylalanine residue, a glycine residue, a glutamic acid residue, an aspartic acid residue, a cysteine residue, a glutamic acid residue, a histidine residue, an isoleucine residue, a leucine residue, a lysine residue, a methionine residue, a proline residue, a serine residue, a threonine residue, a tryptophan residue, a tyrosine residue, or a valine residue; p' is 2-4;
L2is composed of
Figure BDA0002112110080000031
Figure BDA0002112110080000032
Wherein, c terminal and L1E terminal of (a) is connected to the f terminal of (b) and L3Are connected to end d of n2Independently 1 to 8;
L3is composed of
Figure BDA0002112110080000041
Wherein the b terminal is connected to the Ab, and the d terminal is connected to the L2Are connected.
In a preferred embodiment of the invention, the antibody may be an antibody conventional in the field of anti-tumor ADCs, preferably the anti-HER 2 antibody Trastuzumab or a variant thereof, the anti-B7-H3 antibody P2E5 or a variant thereof, the anti-claudin 18.2 antibody IMAB362 or a variant thereof, or the anti-Trop 2 antibody RS7 or a variant thereof, further preferably the anti-HER 2 antibody Trastuzumab or a variant thereof, most preferably the 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 the 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 anti-Claudin18.2 antibody IMAB362 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, L is3The b segment of (a) is preferably linked to a thiol group on the antibody in the form of a thioether bond. To be provided with
Figure BDA0002112110080000042
For the purpose of example only,
Figure BDA0002112110080000043
the connection form with cysteine residue in the antibody is
Figure BDA0002112110080000044
In a preferred embodiment of the present invention, D can be a cytotoxic drug conventional in the ADC field, and the present invention is particularly preferably a cytotoxic drug containing a hydroxyl group or an amino group, more preferably a topoisomerase inhibitor containing a hydroxyl group or an amino group, even more preferably a topoisomerase I inhibitor containing a hydroxyl group or an amino group, even more preferably camptothecin or a derivative thereof, and most preferably camptothecin or a derivative thereof
Figure BDA0002112110080000051
Said L1Preferably, the compound is bonded to a hydroxyl group of the cytotoxic drug in the form of an ether bond or to an amino group of the cytotoxic drug. When said L is1When attached to an amino group of the cytotoxic drug, the hydrogen of the amino group is unsubstituted or substituted with an R3Substitution; said R3Is C1～C6Alkyl radical, C3～C8Cycloalkyl radical, C6～C14Aryl, 5-to 10-membered heteroaryl or-C (═ O) R3-1(ii) a The heteroatom in the 5-10 membered heteroaryl is selected from one or more of N, O and S, and the number of the heteroatoms is 1, 2, 3 or 4; r3-1Is hydroxy-substituted C1～C4An alkyl group. When said L is1After connection with said D, L1D is preferably
Figure BDA0002112110080000052
With L2Is composed of
Figure BDA0002112110080000053
For example, said L1D can be
Figure BDA0002112110080000054
With L2Is composed of
Figure BDA0002112110080000055
Figure BDA0002112110080000056
For example, said L1D can be
Figure BDA0002112110080000057
In a preferred embodiment of the present invention, m is preferably 7 to 8, and is further preferably 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7 or 7.8.
In a preferred embodiment of the invention, when said R is1Is substituted or unsubstituted C1～C10When alkyl, said C1～C10The alkyl group is preferably C1～C4The alkyl group is more preferably a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, an isobutyl group or a tert-butyl group, and most preferably a methyl group or an ethyl group.
In a preferred embodiment of the invention, when said R is1Is substituted or unsubstituted C6～C14When aryl, said C6～C14The aryl group is preferably a phenyl group, a naphthyl group or an anthryl group, and more preferably a phenyl group.
In a preferred embodiment of the present invention, R is1-1、R1-2And R1-3Independently, a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, an isobutyl group, or a tert-butyl group is preferable, and a methyl group is more preferable.
In a preferred embodiment of the invention, R1When substituted C1～C10The substituents in the alkyl radical being hydroxy-substituted C1～C4At alkoxy, said C1～C4The alkoxy group is preferably a methoxy group, an ethoxy group, a n-propoxy group, an isopropoxy group, a n-butoxy group, an isobutoxy group, or a tert-butoxy group, and more preferably an ethoxy group.
In a preferred embodiment of the invention, when said R is3is-C (═ O) R3-1When R is said3-1Preferably, the alkyl group is a hydroxyl-substituted methyl group, a hydroxyl-substituted ethyl group, a hydroxyl-substituted n-propyl group, a hydroxyl-substituted isopropyl group, a hydroxyl-substituted n-butyl group, a hydroxyl-substituted isobutyl group, or a hydroxyl-substituted tert-butyl group, and more preferably a hydroxyl-substituted methyl group.
In a preferred embodiment of the present invention, L is3Preferably, it is
Figure BDA0002112110080000061
In a preferred embodiment of the invention, said L is independently preferably a glycine residue or a phenylalanine residue; said p is preferably 4. Said (L) p is preferably
Figure BDA0002112110080000062
Wherein the g terminus is attached to the carbonyl group in formula I.
In a preferred embodiment of the present invention, R is1Preferably substituted or unsubstituted C1～C10Alkyl, or substituted or unsubstituted C6～C14An aryl group; said substituted C1～C10Alkyl and substituted C6～C14The substituent in the aryl group is preferably one or more groups selected from the group consisting of the following groups, and when a plurality of substituents are present, the substituents are the same or different: -NR1- 1R1-2、-S(O)2R1-3And hydroxy-substituted C1～C4An alkoxy group.
In a preferred embodiment of the present invention, R is2Hydrogen is preferred.
In a preferred embodiment of the present invention, n is1Preferably 2.
In a preferred embodiment of the present 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 preferably
Figure BDA0002112110080000071
Wherein the h-terminus is attached to the carbonyl group in formula II.
In a preferred embodiment of the present invention, said
Figure BDA0002112110080000072
Preferably, it is
Figure BDA0002112110080000073
In a preferred embodiment of the present invention, said
Figure BDA0002112110080000074
Preferably, it is
Figure BDA0002112110080000081
In a preferred embodiment of the present invention, L is2Preferably, it is
Figure BDA0002112110080000082
Figure BDA0002112110080000083
In a preferred embodiment of the present invention, n is2Preferably 1.
In a preferred embodiment of the invention, when L1D is
Figure BDA0002112110080000084
When said L is2Preferably, it is
Figure BDA0002112110080000085
In a preferred embodiment of the present invention, R is3preferably-C (═ O) R3-1。
In a preferred embodiment of the invention, when L1D is
Figure BDA0002112110080000086
Figure BDA0002112110080000087
When the above-mentioned (L ') p' is a group
Figure BDA0002112110080000088
In a preferred embodiment of the present invention, when D is
Figure BDA0002112110080000091
When L is1D is preferably
Figure BDA0002112110080000092
In a preferred embodiment of the invention, in the antibody conjugate, the Ab is anti-HER 2 antibody Trastuzumab; d is a cytotoxic drug; m is 2-8;
said L1The structure of (A) is shown as formula I or II;
Figure BDA0002112110080000093
l is independently phenylalanine residue or glycine residue; p is 0 to 4;
said R1Is substituted or unsubstituted C1～C10Alkyl, or substituted or unsubstituted C6～C14An aryl group; said substituted C1～C10Alkyl and substituted C6～C14The substituent in the aryl group is one or more groups selected from the group consisting of the following groups, and when a plurality of substituents are present, the substituents are the same or different: -NR1-1R1-2、-S(O)2R1-3And hydroxy-substituted C1～C4An alkoxy group; said R1-1R is as described1-2And said R1-3Independently is C1～C4An alkyl group;
said R2Is hydrogen;
n is1Is 2;
l' is independently valine residue, alanine residue, lysine residue, phenylalanine residue or citrulline residue; p' is 2-4;
when said L is1When attached to an amino group of the cytotoxic drug, the hydrogen of the amino group is unsubstituted or substituted with an R3Substitution; said R3is-C (═ O) R3-1；R3-1Is hydroxy-substituted C1～C4An alkyl group;
said L2Is composed of
Figure BDA0002112110080000101
N is21 to 8;
said L3Is composed of
Figure BDA0002112110080000102
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,in the antibody coupling medicament, the Ab is an anti-HER 2 antibody Trastuzumab; d is
Figure BDA0002112110080000103
M is 7-8;
said L1The structure of (A) is shown as formula I or II;
Figure BDA0002112110080000104
l is independently phenylalanine residue or glycine residue; p is 4;
said R1Is substituted or unsubstituted C1～C4Alkyl, or substituted or unsubstituted C6～C14An aryl group; said substituted C1～C4Alkyl and substituted C6～C14The substituent in the aryl group is one or more groups selected from the group consisting of the following groups, and when a plurality of substituents are present, the substituents are the same or different: -N (CH)3)2、-S(O)2CH3And hydroxy-substituted ethoxy;
said R2Is hydrogen;
n is1Is 2;
l' is independently valine residue, alanine residue, lysine residue, phenylalanine residue or citrulline residue; p' is 2-4;
when said L is1When attached to an amino group of the cytotoxic drug, the hydrogen of the amino group is unsubstituted or substituted with an R3Substitution; said R3is-C (═ O) R3-1；R3-1Is hydroxy-substituted C1～C4An alkyl group;
said L2Is composed of
Figure BDA0002112110080000111
N is21 to 8;
said L3Is composed of
Figure BDA0002112110080000112
And when L is1D is
Figure BDA0002112110080000113
When said L is2Is composed of
Figure BDA0002112110080000114
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 invention, in the antibody conjugate, the Ab is anti-HER 2 antibody Trastuzumab; l is1D is
Figure BDA0002112110080000115
m is 7-8;
said L1The structure of (A) is shown as formula I or II;
Figure BDA0002112110080000116
l is independently phenylalanine residue or glycine residue; p is 4;
said R1Is substituted or unsubstituted C1～C4Alkyl, or substituted or unsubstituted C6～C14An aryl group; said substituted C1～C4Alkyl or substituted C6～C14The substituent in the aryl group is one or more groups selected from the group consisting of the following groups, and when a plurality of substituents are present, the substituents are the same or different: -N (CH)3)2、-S(O)2CH3And hydroxy-substituted ethoxy;
said R2Is hydrogen;
n is1Is 2;
l' is independently valine residue, alanine residue, lysine residue, phenylalanine residue or citrulline residue; p' is 2-4;
said R3is-C (═ O) R3-1；R3-1Is a hydroxy-substituted methyl group;
said L2Is composed of
Figure BDA0002112110080000121
N is21 to 8;
said L3Is composed of
Figure BDA0002112110080000122
And when L is1D is
Figure BDA0002112110080000123
When said (L ') p' is
Figure BDA0002112110080000124
When L is1D is
Figure BDA0002112110080000125
When said L is2Is composed of
Figure BDA0002112110080000126
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 represented by any one of the following:
Figure BDA0002112110080000127
Figure BDA0002112110080000131
Figure BDA0002112110080000132
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 the 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 anti-Claudin18.2 antibody IMAB362 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 represented by any one of the following:
Figure BDA0002112110080000141
Figure BDA0002112110080000142
further preferred is
Figure BDA0002112110080000143
Figure BDA0002112110080000151
Figure BDA0002112110080000152
Further preferred is
Figure BDA0002112110080000153
Figure BDA0002112110080000154
Figure BDA0002112110080000155
Wherein Ab is 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 conjugate of the connecting group and the drug, and the structural general formula of the conjugate is L4-L2-L1-D, wherein L4Is composed of
Figure BDA0002112110080000161
L2、L1And D is as previously defined, L2F terminal of (1) and L4Are connected.
In a preferred embodiment of the present invention, the linker-drug conjugate is preferably any one of the compounds shown below:
Figure BDA0002112110080000162
the invention provides a preparation method of the antibody conjugate drug, which comprises the following steps of conjugating the linker-drug conjugate with an antibody.
In the present invention, the conditions and procedures for the coupling may be those conventional in the art.
The invention also provides a pharmaceutical composition, which comprises the antibody coupling drug and a pharmaceutically acceptable carrier.
The invention also provides application of the antibody conjugate medicine or the pharmaceutical composition in preparing a medicine for preventing or treating cancer. The cancer is preferably gastric 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 called DAR, i.e. drug antibody coupling ratio), and m can be an integer or a decimal, preferably understood as: the average value of the molar ratio of the drug molecules to the monoclonal antibody molecules in the antibody conjugate drug obtained by conjugating a single monoclonal antibody molecule to a cytotoxic drug can be generally determined by using Hydrophobic-Interaction Chromatography (HIC), polyacrylamide-SDS gel electrophoresis (SDS-PAGE, electrophoresis), liquid Chromatography-mass spectrometry (LC-MS), and the like.
Antibodies of the invention can 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 amino acid sequence mutants of antibodies, as well as covalent derivatives of the native polypeptide, provided that the biological activity comparable to that of the native polypeptide is retained. Amino acid sequence mutants typically differ from the native amino acid sequence in that one or more amino acids are substituted for one or more amino acids in the native amino acid sequence 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-terminal and/or C-terminal truncation mutants. Typically, the amino acid sequence mutants are at least 70%, 75%, 80%, 85%, 90%, 95%, 98% or more than 99% homologous to the native sequence.
The term "treatment" or its equivalent when used with reference to, for example, cancer, refers to a procedure or process for reducing or eliminating the number of cancer cells in a patient or alleviating the symptoms of cancer. "treating" 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 alleviated. Generally, methods for treating cancer are performed even with a 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 expectations.
The term "pharmaceutically acceptable carrier" refers to any formulation or carrier medium capable of delivering an effective amount of an active agent of the present invention, without interfering with the biological activity of the active agent, and without toxic side effects to the host or patient, and representative carriers include water, oils, vegetables and minerals, cream bases, lotion bases, ointment bases, and the like. These include suspending agents, viscosity enhancers, skin penetration enhancers, and the like. Their preparation is known to those skilled in the cosmetic or topical pharmaceutical field.
The above preferred conditions can be arbitrarily combined to obtain preferred embodiments of the present invention without departing from the common general knowledge in the art.
The reagents and starting materials used in the present invention are commercially available.
The positive progress effects of the invention are as follows: the novel antibody coupling drug can realize the wide application of cytotoxic drugs in the field of ADC and treat tumor patients with drug resistance to microtubule ADC.
Detailed Description
The invention is further illustrated by the following examples, which are not intended to limit the scope of the invention. The experimental methods without specifying specific conditions in the following examples were selected according to the conventional methods and conditions, or according to the commercial instructions.
Preparation example 1 preparation of DS-8201a linker-drug conjugate GGFG-Dxd
The compound GGFG-Dxd (structure below) was synthesized by reference to the known method reported in WO2015146132A1, ESI-MS m/z: 1034.5(M + H) in the form of a,1H-NMR(400MHz,DMSO-d6)8.61(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).
Figure BDA0002112110080000191
EXAMPLE 1 Synthesis of DX01 Compound
Figure BDA0002112110080000192
Step 1 synthesis of compound IIIC:
commercially available compound IIIA (5.58g,10mmol) and commercially available IIIB (3.16g,10mmol) were mixed in 100mL of anhydrous N, N-dimethylformamide, HATU (1.14g, 3.0mmol) and triethylamine (10 mL) were added, and the mixture was reacted at room temperature for 2 h. After the reaction, the solvent was removed by distillation under reduced pressure, and the crude product was purified by silica gel column chromatography [ chloroform: methanol ═ 10:1(v/v) ] purification afforded the title compound IIIC (7.71g, yield 90%), ESI-MS m/z: 857.1(M + H).
Step 2 synthesis of compound IIID:
compound IIIC (7.71g, 9mmol) was dissolved in dichloromethane, and 9mL of trifluoroacetic acid was added to react at room temperature for 0.5 h. After the reaction was completed, the solvent was removed by distillation under the reduced pressure, and the crude product was purified by silica gel column chromatography [ dichloromethane: purification of methanol 10:1(v/v) ] afforded the title compound IIID as the trifluoroacetate salt (5.90g, 90% yield), ESI-MS m/z: 614.9(M + H).
Step 3 synthesis of compound IIIE:
tert-butyl 2-glycolate (2.5g, 18.9mmol) and di (p-nitrophenyl) carbonate (6.3g, 20.8mmol) were mixed and dissolved in 200mL anhydrous DMF, 25mL triethylamine was added and the reaction was carried out at room temperature for 2 hours. After completion of the reaction of the starting materials as monitored by LC-MS, IIID trifluoroacetate salt (7g, 9.6mmol) was added and the reaction was continued for 1 hour. After the reaction, most of the solvent was distilled off under reduced pressure, then 150mL of water was added, extraction was carried out three times with ethyl acetate (100 mL each), the organic phases were combined after liquid separation, washed with saturated brine, dried over anhydrous sodium sulfate and concentrated, and the resulting crude product was subjected to silica gel column chromatography [ dichloromethane: purification of ethyl acetate ═ 10:1(v/v) ] afforded the title compound IIIE (6.8g, 92% yield), ESI-MS m/z: 773.5(M + H).
Step 4 synthesis of compound IIIF:
compound IIIE (7.73g, 10mmol) was dissolved in 150mL of dichloromethane, and 9mL of trifluoroacetic acid was added to react at room temperature for 0.5 h. After the reaction was completed, the solvent was removed by distillation under the reduced pressure, and the crude product was purified by silica gel column chromatography [ dichloromethane: methanol ═ 10:1(v/v) ] purification afforded the title compound IIIF (5.73g, 80% yield), ESI-MS m/z: 717.2(M + H).
Step 5 synthesis of compound IIIG:
compound IIIF (5.73g, 8mmol) was mixed with commercially available Exatecan mesylate (4.5g, 8mmol) in 30mL of anhydrous DMF and HATU (3.8g, 10mmol) and triethylamine 2mL were added and reacted at room temperature for 2 h. After the reaction, the solvent was removed by distillation under reduced pressure, and the crude product was purified by silica gel column chromatography [ chloroform: methanol ═ 10:1(v/v) ] purification afforded the title compound IIIG (7.89g, yield 87%), ESI-MS m/z: 1134.1(M + H).
Step 6 synthesis of compound DX 01:
compound IIIG (1g, 0.929mmol) was dissolved in 20mL of anhydrous DMF and 0.5mL of 1, 8-diazabicycloundec-7-ene was added and reacted at room temperature for 1 hour. After the starting material had reacted, succinimidyl 6- (maleimido) hexanoate (428.5mg, 1.39mmol) was added directly and stirred at room temperature for 1 hour. The solvent was distilled off under reduced pressure, and the crude product was subjected to silica gel column chromatography [ chloroform: methanol ═ 8:1(v/v) ] purification afforded the title compound (0.96g, 73% yield), ESI-MS m/z: 1105.3(M + H).
Example 2
Synthesis of DX02, DX03 and DX04
Figure BDA0002112110080000211
Referring to example 1, compound IIIA was reacted with a commercially available different amino fragment at step 1, and the subsequent steps were the same as in example 1, to finally obtain compound DX02-DX 04.
Figure BDA0002112110080000212
Figure BDA0002112110080000221
Compound DX 02: pale yellow powder, ESI-MS m/z: 1164.3(M + H); compound DX 03: pale yellow powder, ESI-MS m/z: 1199.5(M + H); compound DX 04: pale yellow powder, ESI-MS m/z: 1169.2(M + H).
EXAMPLE 3 Synthesis of DX05, DX06 Compound
Figure BDA0002112110080000222
Step 1 Synthesis of Compound DXD-1
Commercially available compound DXD (1g, 2.03mmol) and tert-butyldimethylsilyl chloride (0.46g, 3.05mmol) 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 to dryness under reduced pressure, and the crude product was purified by column chromatography [ dichloromethane: methanol ═ 50:1(v/v) ] purification afforded the title compound (1.11g, 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 dichloromethane, 0.5 equivalent of triphosgene was added, and then 2 equivalents of p-dimethylaminopyridine were added to react at room temperature for 1 hour. Adding the intermediate V (R represents different substituents respectively corresponding to DX05 and DX06), then reacting for 0.5 hour at room temperature, and adding 10% trifluoroacetic acid for treating for 1 hour to remove silicon protection. Then evaporating the solvent to dryness under reduced pressure, purifying the crude product by column chromatography to obtain intermediate IX, and obtaining the target product (wherein R in DX05 is methyl, R in DX06 is methyl) in the subsequent reaction steps according to example 1
Figure BDA0002112110080000231
). Compound DX 05: light yellow solid, ESI-MS m/z: 1107.3(M + H); compound DX 06: colorless oil, ESI-MS m/z: 1181.2(M + H).
Example 4 Synthesis of Compounds DX07 and DX08
Figure BDA0002112110080000232
Figure BDA0002112110080000241
Intermediates VIII-5 and IX-5 were obtained according to preparation example 1 or example 2, and then treated with 1, 8-diazabicycloundec-7-ene according to step 6 of example 1 and reacted with commercially available starting material B to give the desired product. Compound DX 07: colorless oil, ESI-MS m/z: 1305.5(M + H); compound DX 08: light yellow oil, ESI-MS m/z: 1305.3(M + H).
EXAMPLE 5 Synthesis of DX09, DX10
Figure BDA0002112110080000251
Step 1 synthesis of compound 2:
commercially available compound 1(10g, 26.4mmol) and 4-azidobutyric acid (5.11g, 39.6mmol) were mixed and dissolved in 100mL of anhydrous DMF, EEDQ (13.1g, 52.8mmol) was added, and the reaction was carried out at room temperature for 5 hours. After the reaction was completed, the solvent was removed by distillation under the reduced pressure, and the residue was purified by silica gel column chromatography [ dichloromethane: purification of methanol 10:1(v/v) ] yielded the title compound (11.8g, 91% yield), ESI-MS m/z: 491.3(M + H).
Step 2 synthesis of compound 3:
compound 2(10g, 20.4mmol) was dissolved in 100mL of anhydrous DMF, cooled to 0 deg.C, thionyl chloride (1.8mL, 24.5mmol) was added slowly and the reaction was continued at this temperature for 30 minutes. After the reaction was completed, ice water was slowly added to quench the reaction, a solid was precipitated, filtered, and the filter cake was washed with water, methyl t-butyl ether, and dried under reduced pressure to obtain the title compound (8.6g, yield 83%) ESI-MS m/z: 509.1(M + H).
Step 3 synthesis of compound 4:
compound 3(1g, 1.97mmol) was mixed with commercially available Exatecan mesylate (1.12g, 1.97mmol) and dissolved in 50mL of anhydrous DMF, and triethylamine (1mL) was added and reacted at room temperature for 2 hours. After the reaction was completed, the solvent was removed by distillation under the reduced pressure, and the crude product was purified by silica gel column chromatography [ dichloromethane: methanol ═ 10:1(v/v) ] purification afforded the title compound (1.1g, 63% yield), ESI-MS m/z: 908.1(M + H).
Step 4 synthesis of compound 5:
compound 4(1g, 1.10mmol) was mixed with glycolic acid (83.7mg, 1.10mmol) and dissolved in 20mL of anhydrous DMF, and HATU (836mg, 2.20mmol) and 1mL of triethylamine were added to react at room temperature for 1.5 hours. After the reaction, the solvent was removed by distillation under the reduced pressure, and the residue was purified by silica gel column chromatography [ dichloromethane: purification of methanol 10:1(v/v) ] yielded the title compound (542mg, yield 51%), ESI-MS m/z: 966.4(M + H).
Step 5 synthesis of compound DX 09:
compound 5(500mg, 0.518mmol) and commercial starting material C (128.6mg, 0.518mmol) were dissolved in 10mL of anhydrous DMF and reacted at room temperature for 1 hour with the addition of a catalytic amount of cuprous bromide. After the reaction, the solvent was removed by distillation under reduced pressure, and the residue was purified by silica gel column chromatography [ chloroform: methanol 10:1(v/v) ] purification gave the title compound (546mg, yield 87%), ESI-MS m/z: 1214(M + H).
Synthesis of compound DX10 referring to the synthesis of DX09, compound DX10 was obtained as a pale yellow solid by replacing 5, the starting compound, with ESI-MS m/z: 1116.3(M + H).
EXAMPLE 6 Synthesis of DX11, DX12
Figure BDA0002112110080000261
Referring to example 5, the dipeptide starting material was replaced with the corresponding commercially available dipeptide starting material in the initial step, and the subsequent steps were the same as in example 5. Compound DX 11: light yellow solid, ESI-MS m/z: 1088.1(M + H). Compound DX 12: pale yellow foamy solid, ESI-MS m/z: 1193.3(M + H).
Example 7 general procedure for ADC preparation
anti-HER 2 antibody Trastuzumab (concentration 15mg/ml) was replaced into 50mM PB/1.0mM EDTA buffer (pH 7.0) using a G25 desalting column, 15 equivalents of TECP were added, and stirred at 37 ℃ for 2 hours to completely open the interchain disulfide bonds of the antibody, followed by adjusting the pH of the antibody solution after reduction to 6.0 using phosphoric acid and lowering the temperature of the water bath to 25 ℃ to prepare for the coupling reaction. The linker-drug conjugates prepared in preparation example 1 and examples 1 to 6 were dissolved in DMSO, respectively, 12 equivalents of linker-drug conjugate linker were pipetted therefrom and dropwise added to the reduced antibody solution, and DMSO was added thereto to a final concentration of 10% (v/v), and the reaction was stirred at 25 ℃ for 0.5 hour, and after the reaction was completed, the sample was filtered using a 0.22um membrane. Purifying by using tangential flow ultrafiltration system to remove unconjugated small molecules, wherein the buffer solution is 50mM PB/1.0mM EDTA solution (pH6.0), adding sucrose with final concentration of 6%, and storing in-20 deg.C refrigerator. The absorbance values were measured at 280nm and 370nm, respectively, using the UV method to calculate the DAR values, and the results are shown in the following Table (Table 1). 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.
TABLE 1 DAR values determined by UV method for different antibody-conjugated drugs (ADC)
ADC numbering Antibodies 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
NCI-N87 cells and SK-BR-3 cells which are stably transfected and highly express Her2 are selected as cell strains for in vitro activity detection of the experiment, and the dose-effect conditions of different antibody coupling drugs on cell killing are observed. Initial selection of plate density for each cell: 2X 103cells/hole, and cell cytotoxic activity is measured after 16-24 hours; then, the final concentration of the antibody conjugate drug prepared in example 7 after sample addition is set to 5000nM as the initial concentration, 10 concentrations (4-10 fold dilution) of the 5000-0.006 nM design series are determined, the change of killing (or inhibition) in 96 hours is observed,
Figure BDA0002112110080000283
Luminescent Cell visual Assay chemiluminescent staining, reading fluorescence data and calculating IC 50. From the activity test results, all the ADCs show certain antitumor activity, the activity of part of the ADCs exceeds DS-8201a, the activity of part of the ADCs is equivalent to DS-8201a, and the activity of part of the ADCs is remarkably weaker than that of DS-8201a, and the results are shown in Table 2.
TABLE 2 in vitro cytotoxic Activity of different ADCs
Figure BDA0002112110080000281
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, 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 plus an internal standard (irinotecan as an internal standard substance) and then the amount of released free drug was measured by high performance liquid chromatography, and the results are shown in Table 3.
TABLE 3 stability evaluation of part of ADCs in a weakly acidic Environment
Figure BDA0002112110080000282
Figure BDA0002112110080000291
The stability result shows that the stability of the ADC obtained by adopting the new technical scheme is stronger than that of DS-8201a in a weak acid environment, and various normal tissues in a human body are weak acid, so that the ADC provided by the invention is expected to show better safety and similar effectiveness than that of DS-8201 a.
SEQUENCE LISTING
<110> Shanghai Compound Dangjiang biomedical corporation
<120> antibody coupling drug, intermediate thereof, preparation method and application
<130> P19011374C
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<170> PatentIn version 3.5
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Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn
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Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
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Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
275 280 285
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
290 295 300
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
305 310 315 320
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
325 330 335
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
340 345 350
Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser
355 360 365
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
370 375 380
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
385 390 395 400
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
405 410 415
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
420 425 430
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
435 440 445
Pro Gly Lys
450
<210> 5
<211> 214
<212> PRT
<213> Artificial Sequence
<220>
<223> Trastuzumab light chain sequence
<400> 5
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Val Asn Thr Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Ser Ala Ser Phe Leu Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Arg Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln His Tyr Thr Thr Pro Pro
85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 6
<211> 450
<212> PRT
<213> Artificial Sequence
<220>
<223> Trastuzumab heavy chain sequence
<400> 6
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Asn Ile Lys Asp Thr
20 25 30
Tyr Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Arg Ile Tyr Pro Thr Asn Gly Tyr Thr Arg Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ser Arg Trp Gly Gly Asp Gly Phe Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val
115 120 125
Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala
130 135 140
Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser
145 150 155 160
Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val
165 170 175
Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro
180 185 190
Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys
195 200 205
Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp
210 215 220
Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly
225 230 235 240
Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile
245 250 255
Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu
260 265 270
Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His
275 280 285
Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg
290 295 300
Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys
305 310 315 320
Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu
325 330 335
Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr
340 345 350
Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu
355 360 365
Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp
370 375 380
Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val
385 390 395 400
Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp
405 410 415
Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His
420 425 430
Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro
435 440 445
Gly Lys
450
<210> 7
<211> 217
<212> PRT
<213> Artificial Sequence
<220>
<223> P2E5 light chain sequence
<400> 7
Gln Thr Val Val Thr Gln Glu Pro Ser Phe Ser Val Ser Pro Gly Gly
1 5 10 15
Thr Val Thr Leu Thr Cys Gly Leu Asn Ser Gly Ser Val Ser Thr Ser
20 25 30
Tyr Phe Pro Ser Trp Tyr Gln Gln Thr Pro Gly Gln Ala Pro Arg Thr
35 40 45
Leu Ile Tyr Asn Thr Asn Thr Arg Ser Ser Gly Val Pro Asp Arg Phe
50 55 60
Ser Gly Ser Ile Leu Gly Asn Lys Ala Ala Leu Thr Ile Thr Gly Ala
65 70 75 80
Gln Ala Asp Asp Glu Ser Asp Tyr Tyr Cys Leu Leu Tyr Met Asp Ser
85 90 95
Gly Pro His Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly
100 105 110
Gln Pro Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu
115 120 125
Glu Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe
130 135 140
Tyr Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro Val
145 150 155 160
Lys Ala Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn Lys
165 170 175
Tyr Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser
180 185 190
His Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu
195 200 205
Lys Thr Val Ala Pro Thr Glu Cys Ser
210 215
<210> 8
<211> 449
<212> PRT
<213> Artificial Sequence
<220>
<223> P2E5 heavy chain sequence
<400> 8
Gln Val Thr Leu Lys Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Ser
20 25 30
Tyr Met Thr Trp Val Arg Gln Ala Pro Gly Met Gly Leu Glu Trp Val
35 40 45
Ala Ser Met Lys Pro Asp Gly Ser Val Lys His Tyr Val Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Thr Lys Asn Ser Leu Asp
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ser Ser Tyr Asp Thr Arg Trp Gly Trp Phe Asp Pro Trp Gly Glu Gly
100 105 110
Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
115 120 125
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
130 135 140
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
145 150 155 160
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
165 170 175
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
180 185 190
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
195 200 205
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
210 215 220
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro
225 230 235 240
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
245 250 255
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
260 265 270
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
275 280 285
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val
290 295 300
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
305 310 315 320
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
325 330 335
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
340 345 350
Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr
355 360 365
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
370 375 380
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
385 390 395 400
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
405 410 415
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
420 425 430
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
435 440 445
Lys

Claims (15)

1. An antibody coupling medicine with the structural general formula Ab- (L)3-L2-L1-D)m；
Wherein Ab is an antibody;
d is a cytotoxic drug;
m is 2-8;
L1the structure of (A) is shown as formula I or II; wherein the a-terminus is linked to said cytotoxic agent and the e-terminus is linked to said L2The end c of the first and second terminals are connected,
Figure FDA0002112110070000011
l is independently a phenylalanine residue, a glycine residue, a glutamic acid residue, an aspartic acid residue, a cysteine residue, a glutamic acid residue, a histidine residue, an isoleucine residue, a leucine residue, a lysine residue, a methionine residue, a proline residue, a serine residue, a threonine residue, a tryptophan residue, a tyrosine residue, or a valine residue; p is 0 to 4;
R1is hydrogen, substituted or unsubstituted C1～C10Alkyl, substituted or unsubstituted C6～C14An aryl group, a substituted or unsubstituted 5-to 10-membered heterocyclic group, or a substituted or unsubstituted phenoxy group; said substituted C1～C10Alkyl, substituted C6～C14The 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 the following, and when a plurality of substituents are present, the substituents may be the same or different: halogen, hydroxy, -NR1-1R1-2、-S(O)2R1-3、C1～C4Alkyl radical, C1～C4Alkoxy, hydroxy-substituted C1～C4Alkoxy radical, C3～C8Cycloalkyl, 5-to 8-membered heterocyclic group, C6～C10Aryl 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; r1-1、R1-2And R1-3Independently is C1～C4An alkyl group;
R2independently hydrogen, substituted or unsubstituted C1～C10Alkyl, substituted or unsubstituted C3～C10Cycloalkyl, substituted or unsubstituted C6～C14Aryl, substituted or unsubstituted 5-to 10-membered heteroaryl; said substituted C1～C10Alkyl, substituted C3～C10Cycloalkyl, substituted C6～C14The substituents in the aryl group and the substituted 5-to 10-membered heteroaryl group are one or more groups selected from the group consisting of the following groups, and when a plurality of substituents are present, the substituents are the same or different: halogen, hydroxy, C1～C4Alkyl radical, C1～C4Alkoxy radical, C3～C8Cycloalkyl, 5-to 8-membered heterocyclic group, C6～C10Aryl and 5-to 10-membered heteroaryl; the heteroatoms in the 5-to 8-membered heterocyclic group, the 5-to 10-membered heteroaryl group and the substituted or unsubstituted 5-to 10-membered heterocyclic group are selected from one or more of N, O and S, and the number of the heteroatoms is 1, 2, 3 or 4;
n1is 2, 3 or 4;
l' is independently a phenylalanine residue, a glycine residue, a glutamic acid residue, an aspartic acid residue, a cysteine residue, a glutamic acid residue, a histidine residue, an isoleucine residue, a leucine residue, a lysine residue, a methionine residue, a proline residue, a serine residue, a threonine residue, a tryptophan residue, a tyrosine residue, or a valine residue; p' is 2-4;
L2is composed of
Figure FDA0002112110070000021
Figure FDA0002112110070000022
Wherein, c terminal and L1E terminal of (a) is connected to the f terminal of (b) and L3Are connected to end d of n2Independently 1 to 8;
L3is composed of
Figure FDA0002112110070000023
Wherein the b terminal is connected to the Ab, and the d terminal is connected to the L2Are connected.
2. The antibody-conjugated drug of claim 1,
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 the 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 anti-Claudin18.2 antibody IMAB362 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 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 L3The segment b of (a) and the thiol group of the antibody are connected in a thioether bond manner;
and/or D is a cytotoxic drug containing hydroxyl or amino, preferably containing hydroxyl or aminoThe topoisomerase inhibitor having an amino group is more preferably a topoisomerase I inhibitor having a hydroxyl group or having an amino group, still more preferably camptothecin or a derivative thereof, and most preferably camptothecin
Figure FDA0002112110070000031
When said L is1When attached to an amino group of the cytotoxic drug, the hydrogen of the amino group is unsubstituted or substituted with an R3Substitution; said R3Is C1～C6Alkyl radical, C3～C8Cycloalkyl radical, C6～C14Aryl, 5-to 10-membered heteroaryl or-C (═ O) R3-1(ii) a The heteroatom in the 5-10 membered heteroaryl is selected from one or more of N, O and S, and the number of the heteroatoms is 1, 2, 3 or 4; r3-1Is hydroxy-substituted C1～C4An alkyl group;
and/or m is 7-8, preferably 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7 or 7.8;
and/or, when said R is1Is substituted or unsubstituted C1～C10When alkyl, said C1～C10Alkyl is C1～C4An alkyl group, preferably a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, an isobutyl group, or a tert-butyl group, and more preferably a methyl group or an ethyl group;
and/or, when said R is1Is substituted or unsubstituted C6～C14When aryl, said C6～C14Aryl is phenyl, naphthyl or anthracenyl, preferably phenyl;
and/or, said R1-1、R1-2And R1-3Independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, preferably methyl;
and/or, R1When substituted C1～C10The substituents in the alkyl radical being hydroxy-substituted C1～C4At alkoxy, said C1～C4The alkoxy is methoxy, ethoxy, n-propoxy, isopropoxy or n-butylOxy, isobutoxy or tert-butoxy, preferably ethoxy;
and/or, said L3Is composed of
Figure FDA0002112110070000041
And/or, said L is independently a glycine residue or a phenylalanine residue; p is preferably 4; said (L) p is preferably
Figure FDA0002112110070000042
Wherein the g-terminal is attached to the carbonyl group in formula I;
and/or, said R1Is substituted or unsubstituted C1～C10Alkyl, or substituted or unsubstituted C6～C14An aryl group; said substituted C1～C10Alkyl and substituted C6～C14The substituent in the aryl group is preferably one or more groups selected from the group consisting of the following groups, and when a plurality of substituents are present, the substituents are the same or different: -NR1-1R1-2、-S(O)2R1-3And hydroxy-substituted C1～C4An alkoxy group;
and/or, said R2Is hydrogen;
and/or, said n1Is 2;
and/or, said L' is independently a valine residue, an alanine residue, a lysine residue, a phenylalanine residue, or a citrulline residue; p' is preferably 2; said (L ') p' is preferably
Figure FDA0002112110070000043
Figure FDA0002112110070000044
Wherein the h-terminal is connected with the carbonyl group in the formula II;
and/or, said L2Is composed of
Figure FDA0002112110070000051
And/or, said n2Is 1.
3. The antibody-conjugated drug of claim 2,
said R3is-C (═ O) R3-1(ii) a Said R3-1Preferably a hydroxyl-substituted methyl group, a hydroxyl-substituted ethyl group, a hydroxyl-substituted n-propyl group, a hydroxyl-substituted isopropyl group, a hydroxyl-substituted n-butyl group, a hydroxyl-substituted isobutyl group, or a hydroxyl-substituted tert-butyl group, and more preferably a hydroxyl-substituted methyl group;
and/or when L1D is
Figure FDA0002112110070000052
When said L is2Is composed of
Figure FDA0002112110070000053
And/or when L1D is
Figure FDA0002112110070000054
When said (L ') p' is
Figure FDA0002112110070000055
And/or, when D is
Figure FDA0002112110070000056
When L is1D is preferably
Figure FDA0002112110070000057
4. The antibody-conjugated drug of claim 1,
the Ab is anti-HER 2 antibody Trastuzumab; d is a cytotoxic drug; m is 2-8;
said L1The structure of (A) is shown as formula I or II;
Figure FDA0002112110070000061
l is independently phenylalanine residue or glycine residue; p is 0 to 4;
said R1Is substituted or unsubstituted C1～C10Alkyl, or substituted or unsubstituted C6～C14An aryl group; said substituted C1～C10Alkyl and substituted C6～C14The substituent in the aryl group is one or more groups selected from the group consisting of the following groups, and when a plurality of substituents are present, the substituents are the same or different: -NR1-1R1-2、-S(O)2R1-3And hydroxy-substituted C1～C4An alkoxy group; said R1-1R is as described1-2And said R1-3Independently is C1～C4An alkyl group;
said R2Is hydrogen;
n is1Is 2;
l' is independently valine residue, alanine residue, lysine residue, phenylalanine residue or citrulline residue; p' is 2-4;
when said L is1When attached to an amino group of the cytotoxic drug, the hydrogen of the amino group is unsubstituted or substituted with an R3Substitution; said R3is-C (═ O) R3-1；R3-1Is hydroxy-substituted C1～C4An alkyl group;
said L2Is composed of
Figure FDA0002112110070000062
N is21 to 8;
said L3Is composed of
Figure FDA0002112110070000063
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 conjugate of claim 4,
the Ab is anti-HER 2 antibody Trastuzumab; d is
Figure FDA0002112110070000071
M is 7-8;
said L1The structure of (A) is shown as formula I or II;
Figure FDA0002112110070000072
l is independently phenylalanine residue or glycine residue; p is 4;
said R1Is substituted or unsubstituted C1～C4Alkyl, or substituted or unsubstituted C6～C14An aryl group; said substituted C1～C4Alkyl and substituted C6～C14The substituent in the aryl group is one or more groups selected from the group consisting of the following groups, and when a plurality of substituents are present, the substituents are the same or different: -N (CH)3)2、-S(O)2CH3And hydroxy-substituted ethoxy;
said R2Is hydrogen;
n is1Is 2;
l' is independently valine residue, alanine residue, lysine residue, phenylalanine residue or citrulline residue; p' is 2-4;
when said L is1When attached to an amino group of the cytotoxic drug, the hydrogen of the amino group is unsubstituted or substituted with an R3Substitution; saidR3is-C (═ O) R3-1；R3-1Is hydroxy-substituted C1～C4An alkyl group;
said L2Is composed of
Figure FDA0002112110070000073
N is21 to 8;
said L3Is composed of
Figure FDA0002112110070000074
And when L is1D is
Figure FDA0002112110070000081
When said L is2Is composed of
Figure FDA0002112110070000082
6. The antibody conjugate of claim 5,
the Ab is anti-HER 2 antibody Trastuzumab; l is1D is
Figure FDA0002112110070000083
Figure FDA0002112110070000084
M is 7-8;
said L1The structure of (A) is shown as formula I or II;
Figure FDA0002112110070000085
l is independently phenylalanine residue or glycine residue; p is 4;
said R1Is substituted or unsubstituted C1～C4Alkyl, or substituted or unsubstituted C6～C14An aryl group; said substituted C1～C4Alkyl or substituted C6～C14The substituent in the aryl group is one or more groups selected from the group consisting of the following groups, and when a plurality of substituents are present, the substituents are the same or different: -N (CH)3)2、-S(O)2CH3And hydroxy-substituted ethoxy;
said R2Is hydrogen;
n is1Is 2;
l' is independently valine residue, alanine residue, lysine residue, phenylalanine residue or citrulline residue; p' is 2-4;
said R3is-C (═ O) R3-1；R3-1Is a hydroxy-substituted methyl group;
said L2Is composed of
Figure FDA0002112110070000091
N is21 to 8;
said L3Is composed of
Figure FDA0002112110070000092
And when L is1D is
Figure FDA0002112110070000093
When said (L ') p' is
Figure FDA0002112110070000094
7. The antibody conjugate of claim 1, wherein the antibody conjugate is a compound selected from the group consisting of:
Figure FDA0002112110070000095
Figure FDA0002112110070000101
Figure FDA0002112110070000102
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, 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 the 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 anti-Claudin18.2 antibody IMAB362 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.
8. The antibody conjugate of claim 7, wherein the antibody conjugate is a compound selected from the group consisting of:
Figure FDA0002112110070000103
Figure FDA0002112110070000111
Figure FDA0002112110070000112
further preferred is
Figure FDA0002112110070000113
Figure FDA0002112110070000114
Figure FDA0002112110070000121
Further preferred is
Figure FDA0002112110070000122
Figure FDA0002112110070000123
Figure FDA0002112110070000124
Wherein Ab is anti-HER 2 antibody Trastuzumab.
9. The antibody conjugate of claim 8, wherein the antibody conjugate is a compound selected from the group consisting of:
Figure FDA0002112110070000125
Figure FDA0002112110070000131
Figure FDA0002112110070000132
further preferred is
Figure FDA0002112110070000133
Figure FDA0002112110070000134
Figure FDA0002112110070000135
Further preferred is
Figure FDA0002112110070000141
Figure FDA0002112110070000142
Figure FDA0002112110070000143
Wherein Ab is 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.
10. A linker-drug conjugate with a general structural formula L4-L2-L1-D, wherein L4Is composed of
Figure FDA0002112110070000144
Figure FDA0002112110070000145
L2、L1And D is as defined in any one of claims 1 to 7, L2F terminal of (1) and L4Are connected.
11. The linker-drug conjugate of claim 10, wherein the linker-drug conjugate is any one of the following compounds:
Figure FDA0002112110070000146
Figure FDA0002112110070000151
12. a method for preparing an antibody-conjugated drug according to any one of claims 1 to 9, comprising the step of conjugating the antibody to the linker-drug conjugate according to claim 10 or 11.
13. A pharmaceutical composition comprising an antibody-conjugated drug as claimed in any one of claims 1 to 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 an antibody-conjugated drug as claimed in any one of claims 1 to 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
CN116867789A (en) * 2021-03-31 2023-10-10 上海复旦张江生物医药股份有限公司 Preparation method of irinotecan derivative and intermediate thereof
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WO2023016568A1 (en) * 2021-08-12 2023-02-16 上海才致药成生物科技有限公司 Bispecific recombinant 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
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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
WO2024235128A1 (en) * 2023-05-12 2024-11-21 四川科伦博泰生物医药股份有限公司 Antibody-drug conjugate and preparation method and use thereof
WO2025002310A1 (en) * 2023-06-29 2025-01-02 石药集团巨石生物制药有限公司 Antibody-drug conjugate and use thereof
US12274754B2 (en) 2024-08-30 2025-04-15 Immunome, Inc. Immunoconjugates and methods

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