KR-20260064748-A - Novel IGG Protease and Method of Using the Same

Abstract

Novel immunoglobulin G (IgG) proteases and fusion proteins containing the same are provided herein. IgG proteases are useful for methods of treating diseases mediated wholly or partially by pathogenic immunoglobulin G (IgG) antibodies, the methods of preventing or treating antibody-mediated rejection (AMR) of an organ allograft in organ transplant patients.

Inventors

• 미첼, 스펜서
• 로즈, 사샤
• 베일리-켈로그, 크리스
• 그리스월드, 칼
• 세틀리프, 마리온
• 튜버티, 린다
• 도프만, 아리엘

Assignees

• 인스메드 인코포레이티드

Dates

Publication Date

20260507

Application Date

20240801

Priority Date

20230801

Claims (20)

1. As a polypeptide variant of the amino acid sequence of SEQ ID NO. 2 [IdeS], said polypeptide variant is one at one or more amino acid positions of SEQ ID NO. 2 at S3, F4, A6, E9, I10, R11, Y12, Y18, V46, A47, N48, I54, T57, N59, G60, K61, E92, H93, E104, L120, K123, F125, E126, Y135, T138, S159, T161, N162, T185, D188, F199, K200, E201, N203, G222, L223, V230, N233, N246, A251, N273, A275, D288, Q293, V294, G296 A polypeptide variant comprising the above amino acid mutations, wherein the polypeptide variant comprises an amino acid sequence that is at least about 75% identical to SEQ ID NO. 2.
2. In claim 1, the polypeptide variants are polypeptide variants that do not have an amino acid mutation at the next amino acid position of sequence number 2: K56, C66, H234, D256 and D258.
3. In claim 1 or 2, the polypeptide variant is a polypeptide variant that does not have an amino acid mutation at one or more of the following amino acid positions of sequence number 2: A32, N33, T35, Q36, F41, D84, R88, E91, N102, M106, N117, H118, E170, S195, N197, K213, S245, K250, A261, K286, S306.
4. A polypeptide variant according to claim 1 or 2, comprising an amino acid sequence that is at least about 80% identical to SEQ ID NO. 2.
5. A polypeptide variant according to claim 1 or 2, comprising an amino acid sequence that is at least about 85% identical to SEQ ID NO. 2.
6. A polypeptide variant according to claim 1 or 2, comprising an amino acid sequence that is at least about 90% identical to SEQ ID NO. 2.
7. A polypeptide variant according to claim 1 or 2, comprising an amino acid sequence that is at least about 95% identical to SEQ ID NO. 2.
8. A polypeptide variant according to any one of claims 1 to 7, wherein one or more amino acid mutations are one or more amino acid substitutions.
9. A polypeptide variant according to any one of claims 1 to 7, wherein the one or more amino acid mutations are two or more amino acid mutations.
10. In claim 9, the polypeptide variant wherein the two or more amino acid mutations are two or more amino acid substitutions.
11. A polypeptide variant according to any one of claims 1 to 7, wherein the one or more amino acid mutations are three or more amino acid mutations.
12. In claim 11, the above three or more amino acid mutations are polypeptide variants in which three or more amino acid substitutions.
13. A polypeptide variant according to any one of claims 1 to 7, wherein one or more amino acid mutations are four or more amino acid mutations.
14. In paragraph 13, the above-mentioned four or more amino acid mutations are polypeptide variants in which four or more amino acid substitutions.
15. A polypeptide variant according to any one of claims 1 to 7, wherein one or more amino acid mutations are five or more amino acid mutations.
16. In paragraph 15, the above-mentioned 5 or more amino acid mutations are polypeptide variants in which 5 or more amino acid substitutions are present.
17. A polypeptide variant according to any one of claims 1 to 7, wherein one or more amino acid mutations are six or more amino acid mutations.
18. In paragraph 17, the above six or more amino acid mutations are polypeptide variants in which six or more amino acid substitutions.
19. A polypeptide variant according to any one of claims 1 to 7, wherein one or more amino acid mutations are seven or more amino acid mutations.
20. In paragraph 19, the above-mentioned 7 or more amino acid mutations are polypeptide variants in which 7 or more amino acid substitutions.

Description

Novel IGG Protease and Method of Using the Same Cross-reference with related applications This application claims priority from U.S. Provisional Application No. 63/517,080 filed August 1, 2023, the disclosures of which are incorporated herein by reference in their entirety. Mention of electronic sequence list The contents of the electronic sequence list (INMD_195_01WO_SeqList_ST26.xml; size: 3,051,481 bytes; and creation date: July 29, 2024) are incorporated herein by reference in their entirety. Immunoglobulin G-degradase (IdeS) of Streptococcus pyogene cleaves all subclasses of human IgG at the hinge region with high specificity. IdeS also catalyzes heavy chain cleavage of some IgG subclasses in various animals. Pathogenic IgG antibodies contribute to the pathogenesis of acute and chronic transplant rejection as well as numerous autoimmune diseases. Furthermore, in some cases, existing antibodies against gene therapy vectors limit a patient's ability to receive such therapy. Therefore, effectively removing such antibodies is a significant clinical challenge. Streptococcus pyogenes (S. pyogenes) is a bacterial pathogen that causes common infections such as tonsillitis and streptococcal pharyngitis; as a result, most humans are exposed to IdeS and are likely to possess anti-IdeS antibodies in their bloodstream. IdeS-specific antibodies have been detected in serum samples from randomized human subjects (likely due to previous streptococcal infections) as well as in intravenous immunoglobulin (IV-Ig) preparations, which are IgG preparations extracted from serum pooled from thousands of donors. Even if a subject does not possess IdeS-specific antibodies prior to the initial administration of IdeS, such antibodies are likely to be generated subsequently. Therefore, because IdeS is an immunogenic protein, when it is used as a therapeutic agent, the immune system of the patient receiving IdeS will often react to it. This immune response may be involved in the production of antibodies specific to IdeS. In general, the immune response to IdeS, specifically the production of IdeS-specific anti-drug antibodies (ADAs), can (i) reduce the efficacy of IdeS, for example, due to ADA binding, and/or (ii) lead to undesirable or even harmful complications, such as hyperinflammatory responses triggered by immune complexes of ADA and IdeS. The present invention addresses the need for a novel IgG protease by providing a novel protease and a method for using the same. In one aspect, the present invention provides a variant of IdeS IgG protease (“IdeS variant”). The IdeS IgG protease comprises the amino acid sequence presented in SEQ ID NO. 2. IdeS variants are (i) one or more amino acid mutations at one or more of the following amino acid positions of sequence number 2: S3, F4, A6, E9, I10, R11, Y12, Y18, V46, A47, N48, I54, T57, N59, G60, K61, E92, H93, E104, L120, K123, F125, E126, Y135, T138, S159, T161, N162, T185, D188, F199, K200, E201, N203, G222, L223, V230, N233, N246, A251, N273, A275, D288, Q293, V294, G296; and (ii) contains an amino acid sequence that is at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 97%, or at least about 98% identical to SEQ No. 2. In further embodiments, the IdeS variant is at least about 30% active of IdeS in an IgG protease enzymatic assay. In further embodiments, the IdeS variant is at least about 50%, at least about 75%, at least about 90%, at least about 100%, or at least about 110% active of IdeS in an IgG protease enzymatic assay. In one embodiment, one or more mutations in the IdeS variant comprise about 5 to about 20, about 10 to about 20, about 12 to about 20, or about 15 to about 20 amino acid substitutions. In another embodiment, one or more mutations in the IdeS variant comprise about 10 to about 35, or about 24 to about 29 amino acid substitutions. In one embodiment, the IdeS variant comprises one or more amino acid substitutions as set forth in Table 1 disclosed herein. In a further embodiment, the IdeS variant does not have amino acid mutations at the following amino acid positions of SEQ ID NO. 2: K56, C66, H234, D256, and D258. That is to say, in this embodiment, the amino acids at positions 56, 66, 234, 256, and 258 are wild-type IdeS residues. In additional embodiments or other embodiments, the IdeS variant does not have an amino acid mutation at one or more of the following amino acid positions of sequence number 2: A32, N33, T35, Q36, F41, D84, R88, E91, N102, M106, N117, H118, E170, S195, N197, K213, S245, K250, A261, K286, S306. That is, in this embodiment as well, the amino acid at the aforementioned numbered positions is a wild-type IdeS residue. In one embodiment, the IdeS variant is one of S3N, F4I, A6S, E9I, I10T, R11D and R11T, one of Y12N, Y18K, V46D, A47E, N48G, I54T, T57D, T57K, T57N, T57R, T57L and T57Q, one of N59D, G60S and G60T, one of K61R, E92R, H93Y, E104R, L120T, K123A, F125W, E126S, Y135H