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CN-122005839-A - Eribulin-derived hydrophilic antibody drug conjugate

CN122005839ACN 122005839 ACN122005839 ACN 122005839ACN-122005839-A

Abstract

An antibody drug conjugate, its use and preparation method are provided. In particular to a hydrophilic linker which can meet the connection of drug molecules, and the obtained antibody drug conjugate has better stability and hydrophilicity and good pharmacokinetic property.

Inventors

  • LI HUAN
  • YAO BING
  • BAO BIN
  • FANG WEI
  • ZHANG XIAODAN
  • ZHU QIWEN
  • ZHANG YAOQING
  • WEI MIAOMIAO
  • HUI XIWU
  • DAN MO

Assignees

  • 石药集团巨石生物制药有限公司

Dates

Publication Date
20260512
Application Date
20251023
Priority Date
20241025

Claims (19)

  1. 1. An antibody drug conjugate having the structure of formula (I): A-(L-D) m (I) Wherein a is a targeting ligand selected from the group consisting of an antibody (e.g., monoclonal antibody) or antigen binding fragment, a small molecule ligand, a polypeptide; L is a linker moiety, one end of which is linked to ligand A and one end of which is linked to drug molecule D; d is an amino group-containing drug molecule or a tautomer, stereoisomer, meso, racemate, enantiomer, diastereoisomer, isotopic label covalently linked to the linker moiety L through an amino group in its molecular structure, wherein the amino group-containing drug molecule is M is an integer or fraction of 1 to 12; the linker moiety-L-is represented by the formula: -L 1 -L 2 -L 3 -L 4 -, Wherein L 1 is selected from Wherein represents a thiol linkage to a, and wherein represents a thiol linkage to L 2 ; L 2 is a spacer selected from the group consisting of-L 2a -L 2b -C (O) -, wherein L 2a is selected from the group consisting of-O-, -S-, -CO-, -C 1 -C 8 alkylene-, -C 3 -C 8 cycloalkylene-, -C 6 -C 14 arylene-, -C 6 -C 14 arylene-C 1 -C 8 alkylene- -C 1 -C 8 alkylene-C 3 -C 8 cycloalkylene-, -5-6 membered heteroarylene-C 1 -C 8 alkylene-, a linear or branched heteroarylene of 1-50 (preferably 1-20, more preferably 1-12, most preferably 1-8) carbon atoms, 1-50 (preferably 1-20, more preferably 1-12, most preferably 1-8) C-atoms, said alkylene, cycloalkylene, arylene, heteroalkylene, heteroarylene, heterocyclylene each optionally being independently selected from C 1 -C 6 alkyl, C-6C-atoms heteroalkyl, C 1 -C 6 alkoxy, hydroxy, Substituted by one or more substituents of amino, carboxyl or C 3 -C 8 cycloalkyl, said alkylene, heterocyclylene, heteroarylene, heteroalkyl containing 1-12 (preferably 1-8) heteroatoms, said alkylene, heterocyclylene, heteroarylene, heteroatoms of heteroalkyl being selected from one or more of N, O or S, preferably L 2a is selected from -CH 2 CH 2 CH 2 CH 2 CH 2 -、-CH 2 CH 2 (OCH 2 CH 2 ) p -、-(OCH 2 CH 2 ) p -、-CH 2 (OCH 2 CH 2 ) p -、-CH 2 CH 2 (OCH 2 CH 2 ) p CH 2 -、-CH 2 CH 2 (OCH 2 CH 2 ) p CH 2 CH 2 -、-CH 2 (OCH 2 CH 2 ) p CH 2 CH 2 -、 Wherein p is an integer from 1 to 12 (preferably 2-8), L 2b is selected from the group consisting of-C 1 -C 8 alkylene-, linear or branched heteroalkylene of 1-50 (preferably 1-12) carbon atoms, the heteroatoms of said heteroalkyl, heterocyclylene, heteroarylene, heteroalkylene being selected from one or more of N, O or S, preferably L 2b is selected from the group consisting of-C 1 -C 8 alkylene-, linear or branched heteroalkylene of 1-8) carbon atoms; l 3 is a polypeptide sequence selected from a peptide residue consisting of 2-8 natural or unnatural amino acids, wherein the amino acids are optionally further substituted with one or more substituents independently selected from C 1 -C 6 alkyl, heteroalkyl of 1-6 carbon atoms, C 1 -C 6 alkoxy, hydroxy, amino, carboxy or C 3 -C 8 cycloalkyl, the heteroalkyl containing 1-5 (preferably 1-3) heteroatoms selected from one or more (e.g., two or three) of N, O or S; L 4 is a self-cleaving fragment selected from: Or alternatively Wherein represents an amide bond with the carboxyl group of L 3 and an amino bond with the drug molecule D; R 2 is selected from the group consisting of linear or branched heteroalkyl groups containing 4-50 (preferably 4-24, more preferably 6-24, most preferably 8-24) structural units of-OCH 2 CH 2 -, peptide chains (preferably sarcosine) containing 4-50 (preferably 4-24, more preferably 8-24, most preferably 10-24) natural or unnatural amino acids, linear or branched heteroalkyl groups containing mono-, oligo-or polysaccharides, R 4 and R 5 are each independently selected from the group consisting of hydrogen, C1-C8 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl, halogen, etc., X is absent or present * Y is C 1 -C 6 alkylene or-R 3 -C (O) -, wherein R 3 is C 1 -C 6 alkylene or an alkylene group containing 1 to 24 (preferably 1 to 8) -OCH 2 CH 2 -structural units; n is an integer from 0 to 6; Preferably, R 2 is selected from the following structures: wherein R, s, t, u is each independently selected from integers from 1 to 50, R a is selected from a bond, a C 1 -C 6 alkylene group (preferably a C 1 -C 3 alkylene group, more preferably a methylene group, an ethylene group, an n-propylene group, an isopropylene group), R b is selected from a C 1 -C 3 alkyl group, preferably a methyl group, an ethyl group, an n-propyl group, an isopropyl group; More preferably, R 2 is selected from the following structures: preferably, Y is C 1 -C 6 alkylene or-R 3 -C (O) -, wherein R 3 is C 1 -C 6 alkylene or-C 1 -C 6 alkylene- (OCH 2 CH 2 ) v -, wherein v is selected from integers from 1 to 12; More preferably, Y is C 1 -C 3 alkylene or-R 3 -C (O) -, wherein R 3 is C 1 -C 3 alkylene or-C 1 -C 3 alkylene- (OCH 2 CH 2 ) v -, wherein v is selected from integers from 1 to 6; Most preferably Y is methylene, ethylene, n-propylene, isopropylene or-R 3 -C (O) -, wherein R 3 is methylene- (OCH 2 CH 2 ) v -, ethylene- (OCH 2 CH 2 ) v -, n-propyl- (OCH 2 CH 2 ) v -, isopropyl- (OCH 2 CH 2 ) v -, wherein v is selected from integers of 1-12); further preferably, Y is selected from the group consisting of-CH 2 -(OCH 2 CH 2 ) 4 C (O) -.
  2. 2. The antibody drug conjugate of claim 1 having the structure of formula (I), tautomer, stereoisomer or pharmaceutically acceptable salt thereof, A is selected from antibodies or antigen binding fragments thereof targeting human EGF receptor-2 (HER 2), folate receptor alpha (FR alpha).
  3. 3. The antibody drug conjugate of any one of claims 1-2 having the structure of formula (I) and tautomers, stereoisomers or pharmaceutically acceptable salts thereof, wherein L 2 is selected from the following structures: * Represents a connection to L 1 and represents a connection to L 3 .
  4. 4. An antibody drug conjugate having the structure of formula (I) according to any one of claims 1-3, wherein L 3 is selected from peptide residues formed from 2-8 amino acids selected from phenylalanine (F), glycine (G), valine (V), citrulline (Cit), glutamic acid (E), alanine (a), lysine (K), C 1 -C 6 alkyl substituted lysine (e.g. C 1 -C 3 alkyl substituted lysine, e.g. methyl substituted lysine, ethyl substituted lysine, n-propyl substituted lysine, isopropyl substituted lysine), and pharmaceutically acceptable salts thereof; preferably, L 3 is selected from the following structures: Wherein, represents connection with L 2 , and represents connection with L 4 .
  5. 5. The antibody drug conjugate of any one of claims 1-4 having the structure of formula (I) and tautomers, stereoisomers or pharmaceutically acceptable salts thereof, wherein L 4 is selected from the group consisting of: X is absent or is * Represents a bond to a carbon atom and represents a bond to an oxygen atom; Y is selected from methylene, ethylene, n-propylene, isopropylene or-R 3 -C (O) -, wherein R 3 is methylene- (OCH 2 CH 2 ) v -, ethylene- (OCH 2 CH 2 ) v -, n-propylene- (OCH 2 CH 2 ) v -, isopropylene- (OCH 2 CH 2 ) v ), wherein v is selected from integers from 1-12, preferably Y is selected from-CH 2 -(OCH 2 CH 2 ) 4 C(O)-;R 4 and R 5 are each independently selected from hydrogen, C1-C8 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl, halogen, and the like; R 2 is selected from the group consisting of a linear or branched heteroalkyl group comprising 4-50 (preferably 4-24, more preferably 6-24, most preferably 8-24) structural units of-OCH 2 CH 2 , a peptide chain comprising 4-50 (preferably 4-24, more preferably 8-24, most preferably 10-24) natural or unnatural amino acids, a linear or branched heteroalkyl group comprising a monosaccharide, oligosaccharide or polysaccharide; * Represents an amide bond with the carboxyl group of L 3 , an amino group with the drug molecule D; preferably, L 4 is selected from: More preferably, L 4 is selected from: Wherein r, s, t, u are each independently selected from integers of 1-50 (e.g 1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、31、32、33、34、35、36、37、38、39、40、41、42、43、44、45、46、47、48、49、50).
  6. 6. The antibody drug conjugate of any one of claims 1-5 having the structure of formula (I) and tautomers, stereoisomers or pharmaceutically acceptable salts thereof, wherein L is selected from the following structures: Wherein v is selected from integers of 1-12 (e.g. 1, 2, 3,4, 5, 6, 7,8, 9, 10, 11, 12), denotes linking to the thiol group of antibody a, denotes linking to the amino group of the drug molecule D, and R 2 is selected from linear or branched heteroalkyl groups comprising 4-50 (preferably 4-24, more preferably 6-24, most preferably 8-24) OCH 2 CH 2 -building blocks, peptide chains comprising 4-50 (preferably 4-24, more preferably 8-24, most preferably 10-24) natural or unnatural amino acids, linear or branched heteroalkyl groups comprising monosaccharides, oligosaccharides or polysaccharides.
  7. 7. The antibody drug conjugate having the structure of formula (I) according to any one of claims 1-6, wherein a is selected from antibodies targeting fra, her2 or antigen binding fragments thereof, and tautomers, stereoisomers or pharmaceutically acceptable salts thereof.
  8. 8. The antibody drug conjugate of any one of claims 1-7 having the structure of formula (I), wherein a is selected from an antibody or antigen-binding fragment thereof that targets fra, comprising a heavy chain variable region comprising 3 Complementarity Determining Regions (CDRs) wherein the heavy chain complementarity determining region 1 (HCDR 1) amino acid sequence is shown as SEQ ID NO:03, the heavy chain complementarity determining region 2 (HCDR 2) amino acid sequence is shown as SEQ ID NO:04, the heavy chain complementarity determining region 3 (HCDR 3) amino acid sequence is shown as SEQ ID NO:05, and/or a light chain variable region comprising 3 Complementarity Determining Regions (CDRs) wherein the light chain complementarity determining region 1 (LCDR 1) amino acid sequence is shown as SEQ ID NO:06, the light chain complementarity determining region 2 (LCDR 2) amino acid sequence is shown as SEQ ID NO:07, the light chain complementarity determining region 3 (LCDR 3) amino acid sequence is shown as SEQ ID NO:08, and wherein the amino acid sequence of light chain complementarity determining region 3 (HCDR 3) is shown as SEQ ID NO:08 is numbered according to the rules of the following the specification; Preferably, A is an FR alpha-targeting antibody or antigen-binding fragment thereof, comprising a heavy chain variable region (VH) having the amino acid sequence shown in SEQ ID NO:09 and/or a light chain variable region (VL) having the amino acid sequence shown in SEQ ID NO: 10; preferably, a is an antibody targeting fra, the fra antibody further comprising a heavy chain constant region or variant thereof, and/or a light chain constant region or variant thereof; Or a is an antigen binding fragment targeting fαselected from Fab, fab '-SH, fv, scFv, or F (ab') 2 ; Or a is an antibody or antigen-binding fragment thereof targeting fra, which is a humanized or fully human antibody or antigen-binding fragment thereof; Preferably, A is an FR alpha targeting antibody or antigen binding fragment thereof, said FR alpha targeting antibody comprising a heavy chain and/or a light chain, said heavy chain having an amino acid sequence as shown in SEQ ID NO. 01 and said light chain having an amino acid sequence as shown in SEQ ID NO. 02.
  9. 9. The antibody drug conjugate of any one of claims 1-8 having the structure of formula (I) wherein a is selected from the group consisting of HER 2-targeting antibodies or antigen-binding fragments thereof comprising a heavy chain variable region comprising 3 Complementarity Determining Regions (CDRs) wherein the heavy chain complementarity determining region 1 (HCDR 1) amino acid sequence is shown in SEQ ID NO:15, the heavy chain complementarity determining region 2 (HCDR 2) amino acid sequence is shown in SEQ ID NO:16, the heavy chain complementarity determining region 3 (HCDR 3) amino acid sequence is shown in SEQ ID NO:17, and/or the light chain variable region comprising 3 Complementarity Determining Regions (CDRs) wherein the light chain complementarity determining region 1 (LCDR 1) amino acid sequence is shown in SEQ ID NO:18, the light chain complementarity determining region 2 (LCDR 2) amino acid sequence is shown in SEQ ID NO:19, and the light chain complementarity determining region 3 (LCDR 3) amino acid sequence is shown in SEQ ID NO:20, wherein the amino acid sequence is shown in SEQ ID NO:20, and wherein the numbering is according to the rules; Preferably, A is an HER 2-targeting antibody or antigen-binding fragment thereof, which HER 2-targeting antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) having the amino acid sequence shown in SEQ ID NO. 13 and/or a light chain variable region (VL) having the amino acid sequence shown in SEQ ID NO. 14; Preferably, a is an antibody targeting HER2, the HER 2-targeting antibody further comprising a heavy chain constant region or variant thereof, and/or a light chain constant region or variant thereof; Preferably, a is an antigen binding fragment targeting HER2 selected from Fab, fab '-SH, fv, scFv, or F (ab') 2 ; preferably, a is an antibody or antigen-binding fragment thereof that targets HER2, said HER 2-targeting antibody or antigen-binding fragment thereof being a humanized or fully human antibody or antigen-binding fragment thereof; preferably, A is an HER 2-targeting antibody or antigen binding fragment thereof, the HER 2-targeting antibody comprising a heavy chain having an amino acid sequence shown in SEQ ID NO. 11 and/or a light chain having an amino acid sequence shown in SEQ ID NO. 12.
  10. 10. The antibody drug conjugate of any one of claims 1-9 having the structure of formula (I) and tautomers, stereoisomers or pharmaceutically acceptable salts thereof, wherein a is selected from the group consisting of Trastuzumab (farletuzumab) or Trastuzumab-FcS (farletuzumab-FcS) targeting fra and Trastuzumab (Trastuzumab) or a biological analogue thereof, targeting Her 2.
  11. 11. An antibody drug conjugate having the structure of formula (I) as claimed in any one of claims 1 to 10 and its tautomer, stereoisomer or pharmaceutically acceptable salt selected from the group consisting of the following structures: Wherein m is an integer or fraction of 1 to 12, preferably an integer or fraction of 2-8, more preferably 2, 4, 6, 8, and the antibody is Trastuzumab or Farletuzumab.
  12. 12. A pharmaceutical composition comprising an antibody drug conjugate of formula (I) according to any one of claims 1-11 and its tautomer, stereoisomer or pharmaceutically acceptable salt, and one or more pharmaceutically acceptable excipients.
  13. 13. Use of an antibody drug conjugate having the structure of formula (I) according to any one of claims 1-11, as well as its tautomer, stereoisomer or pharmaceutically acceptable salt, or of a pharmaceutical composition according to claim 12, for the preparation of a medicament for the treatment of cancer.
  14. 14. The use of claim 13, wherein the cancer comprises lung cancer, gastric cancer, oral cancer, ovarian cancer, or the presence of a solid tumor that is positive for expression of fra and/or HER 2.
  15. 15. A linker-drug compound of formula (II) and its tautomers, stereoisomers or pharmaceutically acceptable salts: L’-D(II) Wherein L' is a linker precursor capable of being converted to L of formula (I) thereby forming a covalent bond with a of formula (I); D is an amino group-containing drug molecule, and tautomers, stereoisomers or pharmaceutically acceptable salts thereof, covalently linked to the linker precursor moiety L' by an amino group in the molecular structure thereof, wherein the amino group-containing drug molecule is Wherein the linker precursor moiety L' is represented by the formula: Z-L 1 '-L 2 -L 3 -L 4 -, wherein Z is selected from methylsulfonyl, L 1 ' is selected from Wherein represents a linkage to a methanesulfonyl group, and wherein represents a linkage to L 2 ; l 2 、L 3 、L 4 is as defined in claims 1 to 11.
  16. 16. A linker-drug compound of formula (II) according to claim 15 selected from the group consisting of:
  17. 17. A method for producing an antibody-conjugated drug of formula (I) according to any one of claims 1 to 11, which is produced by reacting a thiol-group-containing antibody with a linker-drug compound of formula (II) according to claim 15 via a thioether.
  18. 18. A process for the preparation of a linker-drug compound of formula (II) according to claim 17, characterized by one or more of the following synthetic routes and processes: Wherein, the Is that One, D' is The definition of L 1 、L 2 、R 2 、D、n、X、Y、R 4 and R 5 is the same as the definition of the corresponding symbols of formula (I) or formula (II) in any one of the preceding claims, the polypeptide formed by two or more amino acids of L 2 '-C (=O) L 2 ,L 3 ' -C (=O) L 3 ;L 3 '-COOH can also be prepared by introducing one or more amino acid fragments in sequence and then reacting with p-nitrophenyl chloroformate, or introducing one or more amino acid fragments first and then reacting with p-nitrophenyl chloroformate, and introducing the remaining one or more amino acid fragments, wherein in the reaction route, the primary amine functional group in L 3 ' or its amino acid fragment can be protected by an amino protecting group selected from Boc, teoc, cbz and the protecting group is removed before reacting with an intermediate containing a carboxylic acid functional group, optionally, a compound of a specific conformation is obtained by a chiral resolution method (such as SFC) in a specific step as required, and then carrying out the subsequent reaction.
  19. 19. A compound of formula (III) and its tautomers or stereoisomers: Wherein, the Is that One of which, L 1 、L 2 、D、n、X、Y、R 4 and R 5 are as defined for the corresponding symbols in formula (I) according to any one of claims 1 to 11 or formula (II) according to any one of claims 15 to 16, L 3 ' -C (=O) -corresponds to the definition of L 3 in formula (I) according to any one of claims 1 to 11.

Description

Eribulin-derived hydrophilic antibody drug conjugate Technical Field The invention belongs to the field of biotechnology and medicine, and particularly relates to two antibody drug conjugates, a preparation method and application thereof. Background Antibody conjugated drugs (ADCs) are devices that deliver drug molecules to tumor cells via linkers to achieve killing of the tumor cells. ADC drugs consist of three parts, antibodies, linkers (Linker) and drug molecules (Payload). The antibody is responsible for targeted recognition of tumor cell surface target antigen, the ADC is enriched in tumor tissue, and after cell internalization, the ADC drug enters cells, and drug molecules (Payload) are released under the dual actions of lysosomes and proteases, so that the tumor cells are killed. There are 19 ADC drugs currently on the market, the Payload of which is auristatins (MMAE, MMAF), maytansinoids (DM 1 and DM 4), calicheamicins, camptothecins, PBD and Duocarmycin, respectively. With the application and popularization of ADC drugs in cancer treatment regimens, drug resistance issues become an important factor limiting their development. The development of novel ADC drugs to improve the efficacy of existing ADC drugs and to solve the subsequent drug resistance problem is becoming a major issue. Eribulin is derived from the anti-tumor natural product halichondrin B (Halichondrin B) with a complex structure. The drug is marketed in the united states in 11 2010, formally enters the domestic market in 2019, and is used for treating metastatic breast cancer and liposarcoma. Eribulin is a targeted microtubule inhibitor capable of inhibiting proliferation of cancer cells by binding tubulin and microtubules. Eribulin has become one of the candidate cytotoxins for ADC drugs due to its high pertinence and killing. In the current clinical study, MORAb-202 of the guard is the first ADC molecule of eribulin, an ADC drug targeting folate receptor alpha (FR alpha). At present, the first-stage clinic is finished, and the second-stage clinic experiment is being performed. The ADC medicine takes Farletuzumab (FR alpha humanized IgG1 monoclonal antibody) as an antibody, takes MC-PEG 2-VC-as a connector, takes Eribulin as cytotoxin, wherein the medicine antibody coupling ratio (DAR) is 4.0. The ADC medicine can not only recognize target antigen by depending on antibody and undergo cell internalization, but also kill tumor cells by ADCC, ADCP and CDC effects. In 2020, when Her2 is used as an antibody, MC-PEG2-VC-Eribulin is linker-payload, an ADC drug targeting Her2 is developed, and clinical primary results show that in 5 patients receiving treatment at a dosage level of 1.0mg/kg, 2 patients achieve Partial Remission (PR) in 5 patients receiving treatment, 2 patients have Stable Disease (SD), the optimal total remission rate (BOR) is 40.0% (2/5), and the Disease Control Rate (DCR) is 80.0% (4/5). In addition, the traditional coupling mode of maleimide and cysteine is adopted in the ADC drugs on the market at present, and a linker is also adopted in the non-hydrophilic linker, so that 2 problems are mainly brought about, namely, 1. The ADC drugs constructed by maleimide are easy to undergo reverse Michael addition, so that off-target effect is caused, and 2. The hydrophobicity of linkder-drug causes the ADC to generate high-proportion polymers and influence PK. The invention provides a linker with a hydrophilic group, which can meet the connection of eribulin, and the obtained ADC has better stability, hydrophilicity and good pharmacokinetic property, and the ADC medicine is expected to solve the medicine resistance problem of the marketed medicine with the same kind of indications. Disclosure of Invention The present invention provides an antibody drug conjugate having the structure of formula (I): A-(L-D)m (I) Wherein a is a targeting ligand selected from the group consisting of an antibody (e.g., monoclonal antibody) or antigen binding fragment, a small molecule ligand, a polypeptide; L is a linker moiety, one end of which is linked to ligand A and one end of which is linked to drug molecule D; d is a drug molecule containing amino groups or a tautomer, stereoisomer or pharmaceutically acceptable salt thereof, covalently linked to the linker moiety L through the amino groups in the molecular structure thereof; m represents the molar ratio of drug molecule to antibody a (also known as DAR, i.e. drug-antibody coupling ratio). M is an integer or fraction of 1 to 12 (e.g., 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12). When m is a fraction, it refers to the average number of linker-drug molecules (L-D) conjugated per targeting ligand unit (a). In some embodiments, a is selected from an antibody or antigen binding fragment thereof that targets HER2 (ErbB 2) or fra. In some embodiments, a is a modification, e.g., an alteration, increase or decrease of one or more amino acids. In some embodiments, a is selected from an antibody or antigen bi