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KR-102964486-B1 - Improved Conjugation Linker

KR102964486B1KR 102964486 B1KR102964486 B1KR 102964486B1KR-102964486-B1

Abstract

A β-removal linker suitable for the conjugation of small molecules, peptides, and proteins, and a compound comprising said linker are provided.

Inventors

  • 애슐리, 게리 더블유.
  • 헌, 브라이언
  • 폰테인, 숀
  • 슈나이더, 에릭 엘.

Assignees

  • 프로린크스 엘엘시

Dates

Publication Date
20260513
Application Date
20200403
Priority Date
20190405

Claims (20)

  1. As a linker drug of chemical formula (II), (II), Here: n is an integer from 0 to 6; R 1 is -CN, -SO 2 N(CH 3 ) 2 , -SO 2 CH 3 , -SO 2 Ph, -SO 2 PhCl, -SO 2 N(CH 2 CH 2 ) 2 O, -SO 2 CH(CH 3 ) 2 , -SO 2 N(CH 3 )(CH 2 CH 3 ), or -SO 2 N(CH 2 CH 2 OCH 3 ) 2 ; R 2 is H and; Each R4 is independently a C1 - C3 alkyl; Z is a functional group for linking a linker-drug to an insoluble matrix (M), wherein Z is selected from the group consisting of amines, aminooxy, ketones, aldehydes, maleimidyl, thiols, alcohols, azides, 1,2,4,6-tetraazinyl, trans-cyclooctenyl, bicyclononinyl, and cyclooctinyl; D is a drug; If D is a drug linked via an amine, Y is absent, or if D is a drug linked via a phenol, alcohol, thiol, thiophenol, imidazole, or non-basic amine, Y is -N( R6 ) CH2- ; where R6 is a linker-drug, which is a C1 - C6 alkyl, aryl, or heteroaryl.
  2. In paragraph 1, Z is a linker drug that is an azide.
  3. In paragraph 1, D is a linker drug that is a peptide drug.
  4. In paragraph 1, R 1 is a linker drug that is -CN.
  5. In paragraph 1, the linker drug in which R 1 is -SO 2 CH 3 .
  6. A linker-drug according to any one of claims 1 to 5, wherein each R 4 is methyl.
  7. As a conjugate of chemical formula (III), (III), Here: n is an integer from 0 to 6; R 1 is -CN, -SO 2 N(CH 3 ) 2 , -SO 2 CH 3 , -SO 2 Ph, -SO 2 PhCl, -SO 2 N(CH 2 CH 2 ) 2 O, -SO 2 CH(CH 3 ) 2 , -SO 2 N(CH 3 )(CH 2 CH 3 ), or -SO 2 N(CH 2 CH 2 OCH 3 ) 2 ; R 2 is H and; Each R4 is independently a C1 - C3 alkyl or two R4s form a 3-6 member ring with the carbon atom to which they are attached; D is a drug; If D is a drug linked via an amine, Y is absent, or if D is a drug linked via a phenol, alcohol, thiol, thiophenol, imidazole, or non-basic amine, Y is -N( R6 ) CH2- ; where R6 is a C1 - C6 alkyl, aryl, or heteroaryl; M is an insoluble matrix; q is the multiplicity; Z* is a conjugate, a functional group coupled to an insoluble matrix (M).
  8. In claim 7, Z* is a conjugate comprising a carboxylamide, an oxime, a 1,2,3-triazole, a thioether, a thiosuccinimide, or an ether.
  9. In paragraph 7, R 1 is a conjugate that is -CN.
  10. In paragraph 7, R1 is a conjugate of -SO2CH3 .
  11. A conjugate according to any one of claims 8 to 10, wherein each R4 is independently a C1 - C3 alkyl.
  12. In paragraph 11, each R 4 is a conjugate that is methyl.
  13. A conjugate comprising an insoluble matrix (M) and a linker drug unit of the following chemical formula, , Here: n is an integer from 0 to 6; R 1 is -CN, -SO 2 N(CH 3 ) 2 , -SO 2 CH 3 , -SO 2 Ph, -SO 2 PhCl, -SO 2 N(CH 2 CH 2 ) 2 O, -SO 2 CH(CH 3 ) 2 , -SO 2 N(CH 3 )(CH 2 CH 3 ), or -SO 2 N(CH 2 CH 2 OCH 3 ) 2 ; R 2 is H and; Each R4 is independently a C1 - C3 alkyl, or two R4s form a 3-6 member ring with the carbon atom to which they are attached; Z* is a functional group coupled to the above-mentioned insoluble matrix (M), and Z* comprises 1,2,3-triazole; D is a drug; If D is a drug linked via an amine, Y is absent, or if D is a drug linked via a phenol, alcohol, thiol, thiophenol, imidazole, or non-basic amine, Y is -N( R6 ) CH2- ; where R6 is a C1 - C6 alkyl, aryl, or heteroaryl; A conjugate representing the attachment point of a linker drug unit to the above-mentioned insoluble matrix (M).
  14. In Paragraph 13, Y is the conjugate that is absent.
  15. In paragraph 14, the drug (D) is a conjugate that is a peptide drug.
  16. In claim 15, the above-mentioned insoluble matrix (M) is a conjugate comprising a polyethylene glycol polymer.
  17. In paragraph 16, the polyethylene glycol polymer is a conjugate that is a multiple-armed polymer.
  18. In paragraph 15, R1 is a conjugate that is -CN, -SO₂N ( CH₃ ) ₂ , -SO₂CH₃ , -SO₂Ph, -SO₂PhCl , -SO₂N ( CH₂CH₂ ) ₂O , -SO₂CH ( CH₃ ) ₂ , -SO₂N ( CH₃ )( CH₂CH₃ ), or -SO₂N ( CH₂CH₂OCH₃ ) ₂ .
  19. In paragraph 15, R1 is a conjugate that is -SO2N ( CH3 ) 2 or -SO2CH3 .
  20. In paragraph 15, each R 4 is independently a C 1 -C 3 alkyl conjugate.

Description

Improved Conjugation Linker Cross-reference of related applications [1] The present application claims priority to U.S. provisional application No. 62/830,280, filed April 15, 2019, the entirety of which is incorporated herein by reference. Inclusion by reference to sequence list [2] The present application is filed with a sequence list in electronic form. The sequence list is provided in a file named 670572002140SeqList.txt, which is 2,112 bytes in size and was created on April 3, 2020. The information in the sequence list in electronic form is incorporated by reference in its entirety into the present application. field [3] The present invention relates to a β-removal linker suitable for conjugation of small molecules, peptides, oligonucleotides, and proteins, and a compound comprising said linker. [4] Drug molecules are covalently bonded to macromolecular carriers to improve pharmacological properties such as half-life, stability, solubility, tolerability, and safety. U.S. Patents 8,680,315, 8,754,190, and 9,649,385 disclose drug conjugate systems having β-elimination linkers that release drugs via a rate-controlled β-elimination mechanism. However, along with the released drug or the cleaved crosslink, the β-elimination process generates linker residues bonded to macromolecular carriers containing alkenyl groups, which can be activated for nucleophilic addition. As illustrated in FIG. 1, potential nucleophiles for such addition under physiological conditions include thiols and amines, for example, those present in significant amounts in proteins, or, more preferably, amines released from crosslink cleavage in hydrogels. Although amines are protonated and are expected to be non-reactive at physiological pH, it has been unexpectedly discovered that such Aza-Michael addition occurs, at least in in vitro settings. Previously disclosed linkers (i.e., U.S. Patents 8,680,315 and 8,754,190) provide means to mitigate this unwanted reaction through the addition of an alkyl group on a carbon having a leaching oxygen (corresponding to the R5 group of Formula (I) in U.S. Patent 8,680,315), as illustrated in FIG. 2. An improved linker is needed that can more effectively suppress unwanted Aza-Michael addition. [5] In one embodiment, the linker of chemical formula (I) (I), A linker is provided, wherein n, R1 , R2 , R4 , X, and Z are as disclosed herein. In some embodiments, the linker is a β-removal linker. In some embodiments, the β-removal linker is suitable for the conjugation of small molecules, peptides, and protein therapeutics. [6] In another aspect, the linker-drug of chemical formula (II) (II), This is provided, wherein n, R1 , R2 , R4 , Y, Z, and D are as disclosed herein. In some embodiments, the linker-drug of formula (II) is prepared by combining the linker of formula (I) with a drug such as a small molecule, peptide, or protein therapeutic. [7] In another aspect, the conjugate of chemical formula (III) (III), A conjugate is provided, wherein n, R1 , R2 , R4 , Y, Z*, and D are as disclosed herein. In some embodiments, the conjugate of formula (III) is a conjugate of drug D releasedly connected to a macromolecular carrier M through a linker of formula (I). [8] In another embodiment, a hydrogel of formula (IV), (IV), This is provided, wherein n, r, R1 , R2 , R4 , W, Z*, P1 , and P2 are as disclosed herein. In some embodiments, the compound of formula (IV) is a degradable cross-linked hydrogel. In some embodiments, the degradable cross-linked hydrogel comprises a residue of the linker of formula (I). [9] In another embodiment, a method for preparing compounds of formulas (I), (II), (III), and (IV) and a method for using the same are provided. In another embodiment, a pharmaceutical composition comprising a conjugate of formula (III) or a hydrogel of formula (IV) is provided. [10] FIG. 1 illustrates the cleavage of a carbamate linker in a conjugate as disclosed in U.S. Patent 9,649,385. The intact conjugate (1) undergoes a pH-dependent beta-elimination reaction, in which the linker is cleaved and a free amine (3) is formed along with the linker residue (2). This product can undergo a subsequent reversible aza-Michael reaction to form a relatively stable re-addition adduct (4). If R is an amine-containing drug or a prodrug, the linker is a drug-releasing linker. If R is a second PEG prepolymer, the linker is part of the crosslinking agent of the PEG hydrogel. [11] Figure 2 illustrates the cleavage of a carbamate linker disclosed in U.S. Patent 9,649,385 in a conjugate where both R5 groups are alkyl. Sterile hindrance by the alkyl groups in R5 is expected to slow down the redouble process. [12] Fig. 3 illustrates the cleavage of a γ-substituted carbamate linker disclosed in this specification in a conjugate. [13] Figure 4 shows the aza-michael addition rates of glycine to vinyl sulfone linkers at various pH values. Prism plots of glycine adduct concentration (mM) versus time (h) for Std, β, and Gem diMe linkers: