Cell-penetrating anti-DNA antibodies and uses thereof inhibit DNA repair

Patent metadata

Field	Value
Publication number	US-9701740-B2
Application number	US-201214009327-A
Country	US
Kind code	B2
Filing date	Apr 2, 2012
Priority date	Apr 1, 2011
Publication date	Jul 11, 2017
Grant date	Jul 11, 2017

Abstract

Antibodies that penetrate cell nuclei and inhibit DNA repair or interfere with DNA metabolism are provided for treatment of cancer (both directly and by sensitizing cancer cells to DNA-damaging treatments) or inhibiting or preventing viral infection, proliferation or metabolism. The method involves treating cells with a composition containing cell-penetrating anti-DNA antibodies or derivatives thereof, alone or in combination with treatment that induces DNA damage such as DNA-damaging chemotherapy or radiation. The impact of the cell-penetrating anti-DNA antibodies or derivatives thereof is potentiated in cancer cells that are deficient in DNA repair, and the cell-penetrating anti-DNA antibodies or derivatives thereof are synthetically lethal to cancer cells with DNA repair deficiencies.

First claim

We claim:

1. A method of inhibiting DNA repair in neoplastic or virally transforming or virally transformed cells in a subject, comprising administering to the neoplastic, virally transforming or virally transformed cells in the subject a pharmaceutical composition comprising an effective amount of anti-DNA unconjugated antibodies or antigen binding fragments thereof to impair a DNA repair pathway, wherein all of the anti-DNA unconjugated antibodies or antigen binding fragments thereof of the pharmaceutical composition consist of one or more cell penetrating unconjugated monospecific anti-DNA antibodies or cell penetrating unconjugated antigen binding fragments thereof,
wherein the cell penetrating unconjugated monospecific anti-DNA antibodies or cell penetrating unconjugated antigen binding fragments thereof consist of monoclonal antibody 3E10 produced by ATCC Accession No. PTA 2439 hybridoma, or a cell penetrating antigen binding fragment or humanized form thereof.

2. The method of claim 1 , wherein the neoplastic cells are cancer cells selected from the group consisting of sarcomas, lymphomas, leukemias, carcinomas and adenocarcinomas, blastomas, germ cell tumors, gliomas, neuroendocrine tumors, melanomas, rhabdoid tumors, embryonal tumors, neuroectodermal tumors, carcinoid tumors, craniopharyngiomas, histiocytomas, medulloepitheliomas, mesotheliomas, multiple myelomas, chronic myeloproliferative disease, primitive neuroectodermal tumors, salivary gland tumors, thymomas, thymic carcinoma, thyroid cancer, and Wilms tumor.

3. The method of claim 1 , wherein the cells are radiation resistant.

4. The method of claim 1 , wherein the cells are resistant to chemotherapy.

5. The method of claim 1 , wherein the cells have intrinsic defective or deficient DNA repair.

6. The method of claim 1 , wherein the virally transforming or virally transformed cells are exposed to or infected with a virus having or causing DNA repair defects or deficiencies, or is dependent on host DNA repair pathways for infection, integration, or replication.

7. The method of claim 6 , wherein the virally transforming or virally transformed cells are exposed to or infected with a lentivirus.

8. The method of claim 1 , wherein the cells have one or more mutations in or abnormal expression of DNA repair genes selected from the group consisting of XRCC1, ADPRT (PARP-1), ADPRTL2, (PARP-2), POLYMERASE BETA, CTPS, MLH1, MSH2, FANCD2, PMS2, p53, p21, PTEN, RPA, RPA1, RPA2, RPA3, XPD, ERCC1, XPF, WS19, RAD51, RAD51b, RAD51C, RAD51D, DMC1, XRCCR, XRCC3, BRCA1, BRCA2, PALB2, RAD52, RAD54, RAD50, MRE11, NB51, WRN, BLM, KU70, KU80, ATM, ATR CHK1, CHK2, FANCA, FANCB, FANCC, FANCD1, FANCD2, FANCE, FANCF, FANCG, FANCC, FANCD1, FANCD2, FANCE, FANCF, FANCG, RAD1, and RAD9.

9. The method of claim 1 , wherein the cells have a defective tumor suppressor gene such as BRCA1 or BRCA2.

10. The method of claim 1 , wherein the cells are part of a hypoxic tumor.

11. The method of claim 1 , wherein the antibodies are administered to the subject in combination with a radiosensitizer.

12. The method of claim 11 , wherein the radiosensitizer is selected from the group consisting of cisplatin, doxorubicin, gemcitabine, 5-fluorouracil, PARP1 inhibitors, histone deacetylase inhibitors, proteasome inhibitors, epidermal growth factor receptor (EGRF) inhibitors, insulin-like growth factor-1 (IGF-1) receptor inhibitors, CHK1 inhibitors, mTOR inhibitors, kinase inhibitors, pentoxifylline, and vinorelbine.

13. The method of claim 1 , further comprising treating the subject with radiation therapy, wherein the cell-penetrating anti-DNA antibodies increase the cells' sensitivity to radiation therapy.

14. The method of claim 1 , further comprising treating the subject with an antineoplastic drug that damages DNA or inhibits DNA repair, wherein the cell-penetrating anti-DNA antibodies increase the cells' sensitivity to the antineoplastic drug.

15. The method of claim 1 , further comprising treating the subject with a therapeutic monoclonal antibody.

16. The method of claim 1 , wherein the cell-penetrating anti-DNA antibodies are derived from a subject with an autoimmune disease.

17. The method of claim 16 , wherein the autoimmune disease is systemic lupus erythematous, or an animal model thereof.

18. The method of claim 1 , wherein the cell-penetrating anti-DNA antibodies or antigen binding fragments thereof consist of a single chain variable fragment of the monoclonal antibody 3E10 produced by ATCC Accession No. PTA 2439 hybridoma or a humanized form thereof.

19. The method of claim 1 , wherein the cell-penetrating anti-DNA antibodies or antigen binding fragments thereof are transported into the nucleus of the cell without the aid of a carrier or conjugate.

20. The method of claim 1 , wherein administering the antibodies or antigen binding fragments thereof in combination with chemotherapy or radiation therapy increases DNA damage in the neoplastic or virally transforming or virally transformed cells caused by the radiation or chemotherapy relative to administering of the radiation or chemotherapy alone.

21. The method of claim 1 , wherein the antibodies or antigen binding fragments thereof are not directly cytotoxic to DNA repair-proficient cells.

22. The method of claim 21 , wherein the antibodies or antigen binding fragments are synthetically lethal to DNA repair-deficient cells.

23. A method of treating a subject with cancer, comprising administering to the subject a pharmaceutical composition comprising an effective amount of anti-DNA antibodies or antigen binding fragments thereof to impair a DNA repair pathway in cancer cells in the subject, wherein all of the anti-DNA unconjugated antibodies or antigen binding fragments thereof of the pharmaceutical composition consist of one or more cell penetrating unconjugated monospecific anti-DNA antibodies or cell penetrating unconjugated antigen binding fragments thereof,
wherein the cell penetrating unconjugated monospecific anti-DNA antibodies or antigen binding fragments thereof consist of monoclonal antibody 3E10 produced by ATCC Accession No. PTA 2439 hybridoma, or a cell penetrating unconjugated antigen binding fragments or humanized form thereof.

24. The method of claim 23 , further comprising administering to the subject a DNA damaging agent.

25. The method of claim 24 , wherein the DNA damaging agent is a chemotherapeutic drug or radiation.

26. The method of claim 23 , wherein the subject comprises cancer cells that are DNA repair-deficient.

27. The method of claim 26 , wherein the DNA repair-deficient cells are BRCA1 or BRCA2 deficient.

28. The method of claim 23 , wherein the cell-penetrating anti-DNA antibodies or antigen binding fragments thereof consist of a single chain variable fragment of the monoclonal antibody 3E10 produced by ATCC Accession No. PTA 2439 hybridoma or a humanized form thereof.

29. The method of claim 1 , wherein the pharmaceutical composition comprises an effective amount of the unconjugated cell-penetrating anti-DNA antibodies or antigen binding fragments thereof to reduce DNA repair.

30. The method of claim 23 , wherein the pharmaceutical composition comprises an effective amount of the unconjugated cell-penetrating anti-DNA antibodies or antigen binding fragments thereof to reduce DNA repair.

31. The met

Assignees

• Hansen James E
• Glazer Peter M
• Weisbart Richard H
• Nishimura Robert N
• Chan Grace
• Univ Yale
• Univ California
• The United States Of America Represented By The Dept Of Veterans Affairs

Inventors

• Hansen James E
• Glazer Peter M
• Weisbart Richard H
• Nishimura Robert N
• Chan Grace

Classifications

• C07K16/44

  against material not provided for elsewhere {, e.g. haptens, metals, DNA, RNA, amino acids} · CPC title

• A61K31/513

  having oxo groups directly attached to the heterocyclic ring, e.g. cytosine · CPC title

• A61K31/337

  having four-membered rings, e.g. taxol · CPC title

• A61K45/06

  Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca · CPC title

• A61K39/39558

  against tumor tissues, cells, antigens · CPC title