HUMANIZED ANTIBODIES AND COMPOSITIONS FOR BINDING SPHINGOSINE-1-PHOSPHATE

• US 20090130099A1
• Filed: 10/24/2008
• Published: 05/21/2009
• Est. Priority Date: 10/27/2006
• Status: Active Grant

• Current Assignee: Apollo Endosurgery, Inc. (Boston Scientific Corporation)
• Original Assignee: Lpath Inc. (Boston Scientific Corporation)
• Inventors: HANSEN, Genevieve, SABBADINI, Roger A., JONES, Steven Tarran, GARLAND, William A., WILLIAMS, David
• Granted Patent: US 8,222,373 B2
• Time in Patent Office: Days
• US Class Current: 424/133.100
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 11/06 Antiasthmatics
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: A61P 27/06 Antiglaucoma agents or miotics
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: A61P 35/04 specific for metastasis
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/04 Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C07K 16/44 against material not provided for elsewhere , e.g. haptens, metals, DNA, RNA, amino acids
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/56 : variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/565 : Complementarity determining region [CDR]

Abstract

The present invention relates to anti-S1P agents, particularly humanized monoclonal antibodies (and antigen binding fragments thereof) specifically reactive with S1P, compositions containing such antibodies (or fragments), and the use of such antibodies (or fragments), for example, to treat diseases and conditions associated with aberrant levels of S1P.

Claims

1. An isolated humanized antibody, or an antigen binding fragment thereof, specifically reactive with S1P and comprising at least one heavy chain and at least one light chain, wherein:

A. each heavy chain comprises a heavy chain variable domain comprising;

(i) a sequence of amino acid residues having an amino acid sequence EVQLVQSGAEVKKPGESLKISCQSFGYIFIDHTIHWMRQMPGQGLEWM GAISPRHDITKYNEMFRGQVTISADKSSSTAYLQWSSLKASDTAMYFCA RGGFYGSTIWFDFWGQGTMVTVSS (SEQ ID NO;

32, residues 20-140, inclusive);

or

(ii) a sequence of amino acid residues having at least about 80% sequence identity to the amino acid sequence of A(i), above; and

B. each light chain comprises a light chain variable domain comprising;

(i) a sequence of amino acid residues having an amino acid sequence ETTVTQSPSFLSASVGDRVTITCITTTDIDDDMNWFQQEPG KAPKLLISEGNILRPGVPSRFSSSGYGTDFTLTISKLQPEDF ATYYCLQSDNLPFTFGQGTKLEIK (SEQ ID NO;

33, residues 21-127, inclusive);

or

(ii) a sequence of amino acid residues having at least about 80% sequence identity to the amino acid sequence of B(i), above.

2. An isolated humanized antibody, or an antigen binding fragment thereof, according to claim 1, wherein:

A. each heavy chain comprises at least one of the following;

(i) a first sequence of amino acid residues of sequence DHTIH (SEQ ID NO;

     13);

(ii) a second sequence of amino acid residues AISPRHDITKYNEMFRG (SEQ ID NO;

     31);

or

(iii) a third sequence of amino acid residues of sequence GGFYGSTIWFDF (SEQ ID NO;

     15); and

B. each light chain comprises at least one of the following;

(i) a first sequence of amino acid residues of sequence ITTTDIDDDMN (SEQ ID NO;

     10);

(ii) a second sequence of amino acid residues of sequence EGNILRP (SEQ ID NO;

     11);

or

(iii) a third sequence of amino acid residues of sequence LQSDNLPFT (SEQ ID NO;

     12).

3. An isolated humanized antibody, or an antigen binding fragment thereof, according to claim 1, wherein:

A. each heavy chain comprises;

(i) a first sequence of amino acid residues of sequence DHTIH (SEQ ID NO;

     13);

(ii) a second sequence of amino acid residues AISPRHDITKYNEMFRG (SEQ ID NO;

     31); and

(iii) a third sequence of amino acid residues of sequence GGFYGSTIWFDF (SEQ ID NO;

15); and

B. each light chain comprises;

(i) a first sequence of amino acid residues of sequence ITTTDIDDDMN (SEQ ID NO;

     10);

(ii) a second sequence of amino acid residues of sequence EGNILRP (SEQ ID NO;

     11); and

(iii) a third sequence of amino acid residues of sequence LQSDNLPFT (SEQ ID NO;

     12).

4. An isolated humanized antibody, or an antigen binding fragment thereof, according to claim 1, wherein:

A. each heavy chain comprises a sequence of amino acid residues having an amino acid sequence;

(i) EVQLVQSGAEVKKPGESLKISCQSFGYIFIDHTIHWMRQMPGQGLE WMGAISPRHDITKYNEMFRGQVTISADKSSSTAYLQWSSLKASDTAMY FCARGGFYGSTIWFDFWGQGTMVTVSSASTKGPSVFPLAPSSKSTSG GTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV VTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEL LGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGV EVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPA PIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVE WESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV MHEALHNHYTQKSLSLSPGK (SEQ ID NO;

     38);

or

(ii) a sequence of amino acid residues having at least about 80% sequence identity to the amino acid sequence of A(i), above; and

B. each light chain comprises a sequence of amino acid residues having an amino acid sequence;

(i) ETTVTQSPSFLSASVGDRVTITCITTTDIDDDMNWFQQEPGKAPKL LISEGNILRPGVPSRFSSSGYGTDFTLTISKLQPEDFATYYCLQSDNLPF TFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASWCLLNNFYPREAKV QWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYAC EVTHQGLSSPVTKSFNRGEC (SEQ ID NO;

37, residues 21-234, inclusive);

or

(ii) a sequence of amino acid residues having at least about 80% sequence identity to the amino acid sequence of B(i), above.

5. An isolated humanized antibody, or an antigen binding fragment thereof, according to claim 4, wherein:

A. each heavy chain comprises at least one of the following;

(i) a first sequence of amino acid residues of sequence DHTIH (SEQ ID NO;

     13);

(ii) a second sequence of amino acid residues AISPRHDITKYNEMFRG (SEQ ID NO;

     31);

or

(iii) a third sequence of amino acid residues of sequence GGFYGSTIWFDF (SEQ ID NO;

     15); and

B. each light chain comprises at least one of the following;

(i) a first sequence of amino acid residues of sequence ITTTDIDDDMN (SEQ ID NO;

     10);

(ii) a second sequence of amino acid residues of sequence EGNILRP (SEQ ID NO;

     11);

or

(iii) a third sequence of amino acid residues of sequence LQSDNLPFT (SEQ ID NO;

     12).

6. An isolated humanized antibody, or an antigen binding fragment thereof, according to claim 4, wherein:

A. each heavy chain sequence comprises;

(i) a first sequence of amino acid residues of sequence DHTIH (SEQ ID NO;

     13);

(ii) a second sequence of amino acid residues AISPRHDITKYNEMFRG (SEQ ID NO;

     31); and

(iii) a third sequence of amino acid residues of sequence GGFYGSTIWFDF (SEQ ID NO;

     15); and

B. each light chain sequence comprises;

(i) a first sequence of amino acid residues of sequence ITTTDIDDDMN (SEQ ID NO;

     10);

(ii) a second sequence of amino acid residues of sequence EGNILRP (SEQ ID NO;

     11); and

(iii) a third sequence of amino acid residues of sequence LQSDNLPFT (SEQ ID NO;

     12).

7. An isolated humanized antibody according to claim 1, wherein:

A. each heavy chain comprises a sequence of amino acid residues having an amino acid sequence EVQLVQSGAEVKKPGESLKISCQSFGYIFIDHTIHWMRQMPGQGLE WMGAISPRHDITKYNEMFRGQVTISADKSSSTAYLQWSSLKASDTAMYFCARGGFYGS TIWFDFWGQGTMVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVDKRV EPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVK FNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAP IEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO;

     38); and

B. each light chain comprises a sequence of amino acid residues having an amino acid sequence ETTVTQSPSFLSASVGDRVTITCITTTDIDDDMNWFQQEPGKAPKL LISEGNILRPGVPSRFSSSGYGTDFTLTISKLQPEDFATYYCLQSDNLPFTFGQGTK LEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSG NSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNR GEC (SEQ ID NO;

37, residues 21-234, inclusive).

8. An isolated humanized antibody according to claim 4, wherein each heavy chain comprises a sequence of amino acid residues having an amino acid sequence EVQLVQSGAEVKKPG ESLKISCQSFGYIFIDHTIHWMRQMPGQGLEWMGAISPRHDITKYNEMFRGQVTISADKSSSTA YLQWSSLKASDTAMYFCARGGFYGSTIWFDFWGQGTMVTVSSASTKGPSVFPLAPSSKSTSG GTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVW DVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRSVLTVLHQDWLNGKEYKCKVSN KALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPEN NYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO:

38).

9. An isolated humanized antibody, or an antigen binding fragment thereof, according to claim 1, wherein:

A. each heavy chain comprises the same heavy chain variable domain amino acid sequence as the heavy chain variable domain encoded by the heavy chain gene in vector pATH1009 in ATCC Accession No. PTA-8421; and

B. each light chain comprises the same light chain variable domain amino acid sequence as the light chain variable domain encoded by the light chain gene in vector pATH1009 in ATCC Accession No. PTA-8421.

10. An isolated humanized antibody, or an antigen binding fragment thereof, according to claim 1 wherein at least one amino acid residue of the antibody or antigen binding fragment is glycosylated.

11. An isolated humanized antibody according to claim 1.

12. An isolated humanized antibody according to claim 1 that comprises two heavy chains and two light chains.

13. An isolated humanized antibody according to claim 11, obtainable from CHO cell line LH1 275, as deposited under accession number ATCC PTA-8422.

14. An isolated humanized antibody according to claim 1 as produced in mammalian cells transfected with the plasmid pATH1009 obtainable from E. coli StB12, as deposited under accession number ATCC PTA-8421.

15. An isolated humanized antibody, or an antigen binding fragment thereof, according to claim 1, wherein:

(A) each heavy chain comprises the same heavy chain amino acid sequence as the heavy chain of the antibody expressed by CHO cell line LH1 275, as deposited under accession number ATCC PTA-8422; and

(B) each light chain comprises the same light chain amino acid sequence as the light chain of the antibody expressed by CHO cell line LH1 275, as deposited under accession number ATCC PTA-8422.

16. A pharmaceutical composition comprising an isolated humanized antibody, or an antigen binding fragment thereof, according to claim 1 and a pharmaceutically acceptable carrier.