Chemotherapeutic methods and compositions

• US 9,289,447 B2
• Filed: 09/14/2012
• Issued: 03/22/2016
• Est. Priority Date: 01/06/2009
• Status: Active Grant

• Current Assignee: Gilead Biologics, Inc. (Gilead Sciences Inc.)
• Original Assignee: Gilead Biologics, Inc. (Gilead Sciences Inc.)
• Inventors: Smith, Victoria, Holzer, Alison Kay
• Primary Examiner(s): Xiao, Yan
• Application Number:
  US13/619,139
• Publication Number:
  US 20130017207A1
• Time in Patent Office: 1,285 Days
• Field of Search: 424/146.1, 424/141.1, 424/130.1, 530/350
• US Class Current: 1/1
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 31/282 Platinum compounds
• CPC Class Codes: A61K 31/337 having four-membered rings, e.g. taxol
• CPC Class Codes: A61K 31/513 having oxo groups directly attached to the heterocyclic ring, e.g. cytosine
• CPC Class Codes: A61K 31/517 ortho- or peri-condensed with carbocyclic ring systems, e.g. quinazoline, perimidine
• CPC Class Codes: A61K 31/519 ortho- or peri-condensed with heterocyclic rings
• CPC Class Codes: A61K 31/704 attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin digitoxin A61K31/7048
• CPC Class Codes: A61K 31/7068 having oxo groups directly attached to the pyrimidine ring, e.g. cytidine, cytidylic acid
• CPC Class Codes: A61K 31/7105 Natural ribonucleic acids, i.e. containing only riboses attached to adenine, guanine, cytosine or uracil and having 3'-5' phosphodiester links
• CPC Class Codes: A61K 33/24 Heavy metals; Compounds thereof
• CPC Class Codes: A61K 33/243 Platinum; Compounds thereof
• CPC Class Codes: A61K 39/3955 against proteinaceous materials, e.g. enzymes, hormones, lymphokines
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 16/40 against enzymes
• CPC Class Codes: C12N 15/1137 against enzymes viral enzymes C12N15/1131; receptors C12N15/1138
• CPC Class Codes: C12N 2310/14 interfering N.A.

Abstract

Disclosed herein are methods and compositions for enhancing the cell-killing activity of anti-neoplastic agents by inhibiting the activity of a lysyl oxidase-type enzyme. Also disclosed are methods for screening for chemotherapeutic agents, and for molecules that enhance the activity of chemotherapeutic agents, using cells grown on an extracellular matrix.

Claims

1. A method for killing a malignant cell, the method comprising:

contacting the malignant cell, wherein the malignant cell expresses a lysyl oxidase-type enzyme, with an anti-neoplastic agent and an inhibitor of the lysyl oxidase-type enzyme, wherein the cell is growing in the presence of an extracellular matrix containing collagen, whereby the malignant cell is killed, wherein the anti-neoplastic agent is a chemotherapeutic agent and the inhibitor is an isolated lysyl oxidase-like 2 (LOXL2) antibody or antigen-binding fragment thereof specifically binds to an epitope within the amino acid sequence of SEQ ID NO;

28;

wherein the antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO;

14 and a light chain variable region comprising the amino acid sequence of SEQ ID NO;

15.

3. The method of claim 1, wherein the malignant cell is present in a tumor.

4. The method of claim 1, wherein the chemotherapeutic agent is selected from the group consisting of methotrexate, cispiatin, doxorubicin, docetaxel, erlotinih, paciitaxel, paraclitaxel, 5-fluorouracil and germcitibine.

5. The method of claim 1, wherein the collagen comprises type 1 collagen.

6. The method of claim 1, wherein the LOXL2 antibody or antigen-binding fragment thereof is a Fv, a scFv, a Fab, or a F(ab′

)₂.

7. The method of claim 1, wherein the LOXL2 inhibitor inhibits LOXL2 enzymatic activity.

8. The method of claim 1, wherein the LOXL2 antibody or antigen-binding fragment thereof is polyclonal.

9. The method of claim 1, wherein the antibody or antigen-binding fragment thereof is monoclonal.

10. The method of claim 9, wherein the antibody or antigen-binding fragment thereof is humanized.

11. The method of claim 1, wherein the antibody or antigen-binding fragment thereof comprises the complementarity determining regions CDRs), CDR1, CDR2, and CDR3, of a heavy chain variable region comprising the amino acid sequence of SEQ ID NO;

14, and the CDRs, CDR1, CDR2, and CDR3, of a light chain variable region comprising the amino acid sequence of SEQ ID NO;

15.

12. The method of claim 11, wherein the antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEC) ID NO:

14.

13. The method of claim 11, wherein the antibody or antigen-binding fragment thereof comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO;

15.

14. The method of claim 1, wherein the malignant cell is selected from the group consisting of fibrosarcoma, breast carcinoma, breast adenocarcinoma, pancreatic adenocarcinoma, pancreatic tumor, colon adenocarcinoma, ovarian adenocarcinoma, and lung carcinoma.

2. The method of claim herein the malignant cell is in culture.