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(12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT)

(19) World Intellectual Property Organization International Bureau

(43) International Publication Date

(10) International Publication Number

22 April 2010 (22.04.2010)
(51) International Patent Classification: C07K 16/28 (2006.01) A61P 31/12 (2006.01) A61P 31/04 (2006.01) (21) International Application Number: (22) (25) (26)
(30)

WO 2010/043650 A 3
(74) Agents: BACHELIN, Martin et al; (Hoffmann Arabellastrasse 4 , 81925 Munchen (DE).
Eitle),

(71)

(72) (75)

(81) Designated States (unless otherwise indicated, for every kind of national protection available): AE, AG, AL, AM, PCT/EP2009/06341 1 AO, AT, AU, AZ, BA, BB, BG, BH, BR, BW, BY, BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DK, DM, DO, International Filing Date: DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, 14 October 2009 (14.10.2009) HN, HR, HU, ID, IL, IN, IS, JP, KE, KG, KM, KN, KP, KR, KZ, LA, LC, LK, LR, LS, LT, LU, LY, MA, MD, English Filing Language: ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, English Publication Language: NO, NZ, OM, PE, PG, PH, PL, PT, RO, RS, RU, SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TJ, TM, TN, TR, TT, Priority Data: TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW. 14 October 2008 (14.10.2008) 61/105,259 US Applicant (for all designated States except US): ABL- (84) Designated States (unless otherwise indicated, for every kind of regional protection available): ARIPO (BW, GH, YNX NV [BE/BE]; Technologiepark 4 , B-9052 GhentGM, KE, LS, MW, MZ, NA, SD, SL, SZ, TZ, UG, ZM, Zwijnaarde (BE). ZW), Eurasian (AM, AZ, BY, KG, KZ, MD, RU, TJ, Inventors; and TM), European (AT, BE, BG, CH, CY, CZ, DE, DK, EE, Inventors/ Applicants (for US only): BLANCHETOT, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, LV, Cristoph [FR/NL]; Rhijnvisfeithstraat 9 , NL-2806 R G MC, MK, MT, NL, NO, PL, PT, RO, SE, SI, SK, SM, Gouda (NL). SMIT, Martine [NITNL]; Professor Hans TR), OAPI (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, Frankfurthersingel 236, NL- 1060 T P Amsterdam (NL). ML, MR, NE, SN, TD, TG). LEURS, Regorius [NL/NL]; Saxen Weimarlaan 22-3, NL-1075 C B Amsterdam (NL). JAHNICHEN, Sven Published: [DE/NL]; Geertruida van Lierstaat 70, NL-1069 P E A m s — with international search report (Art. 21(3)) terdam (NL). SCHOLS, Dominique [BE/BE]; Keierveld — before the expiration o the time limit for amending the f 9 , B-3020 Herent (BE). SAUNDERS, Michael John claims and to be republished in the event of receipt of Scott [GB/BE]; Avenue de Ia Jonction 38, B-1 190 B r u s amendments (Rule 48.2(h)) sels (BE). D E HAARD, Johannes Joseph Wilhelmus [NL/NL]; τ Zwint 1, NL-4436 N A Oudelande (NL). — with sequence listing part of description (Rule 5 . 2 (a)) t VANLANDSCHOOT, Peter [BE/BE]; Markettestraat (88) Date of publication of the international search report: 20-A, B-9881 Bellem (BE). VERHEESEN, Peter 13 January 20 11 [NL/BE]; Zeugsteeg 2 , B-9000 Gent (BE).

(54) Title: AMINO ACID SEQUENCES TARGETING HUMAN CD4, CXCR4, CCR5, TLR4, ALPHAV INTEGRIN, BETA3INTEGRIN, BETAL -INTEGRIN, HUMAN ALPHA2- INTEGRIN, CD81, SR-BI, CLAUDIN-I, CLAUDIN-6 AND CLAUDIN-9, RESPECTIVELY, AND NEUTRALIZING VIRAL ENTRY (57) Abstract: The present invention relates t o amino acid sequences that are directed against (as defined herein) human cellular receptors for viruses and/or bacteria such a s e.g. Nanobodies specifically recognizing hCD4, hCXCR4, hCCR5. hTLR4, human alphaV integrin, human beta3 integrin, human betal integrin, human alpha2 integrin, hCD81, hSR-Bl, hClaudin-1, hClaudin-6 and hClaudin-9, a s well a s t o compounds or constructs, and in particular proteins and polypeptides, that comprise or essentially c on sist o f one or more such amino acid sequences. Said amino acid sequences may b e used t o prevent human cell entry o f HIV, HCV, adenoviruses, hantavirus, herpesvirus, echo-virus 1 and others.

A . CLASSIFICATION O F SUBJECT MATTER

INV.

C07K16/28

A61P31/04

A61P31/12

According to International Patent Classification (IPC) o r to both national classification and IPC

B. FIELDS SEARCHED
Minimum documentation searched (classification system followed by classification symbols)

C07K

Documentation searched other than minimum documentation to the extent that such documents are included in the fields searched

Electronic data base consulted dunng the international search (name o f data base and, where practical, search terms used)

EPO-Internal , EMBASE, WPI Data

C . DOCUMENTS CONSIDERED T O B E RELEVANT
Category
*

Citation of document, with indication, where appropriate, of the relevant passages

Relevant t o claim No

WAYNE A MARASCO & JIANHUA SUI: "The growth and potential o f human antiviral monoclonal antibody therapeutics" NATURE BIOTECHNOLOGY, NATURE PUBLISHING GROUP, NEW YORK, NY, U S LNKDD0I:10.1038/NBT1363, vol. 25, 1 December 2007 (2007-12-01), pages 1421-1434, XP009093629 ISSN: 1087-0156 the whole document -/--

1-5,7-9, 16-18, 20-23,25

Further documents are listed i n the continuation of Box C
*

See patent family annex
"T" later document published after the international filing date o r p πo πt y date and not i n conflict with the application but cited t o understand the principle o r theory underlying the invention

Special categones of cited documents

"A" document defining the general state of the art which is not considered t o b e of particular relevance "E" earlier document but published o n o r after the international filing date "L" document which may throw doubts o n p πo πty cla ιm(s) o r which is cited to establish the publication date of another citation o r other special reason (as specified) "O" document refer πng t o a n oral disclosure, use, exhibition o r other means "P" document published prior to the international filing date but later than the priority date claimed Date of the actual completion of the international search

"X" document of particular relevance, the claimed invention cannot be considered novel o r cannot be considered t o involve a n inventive step when the document is taken alone "Y" document o f particular relevance, the claimed invention cannot be considered t o involve a n inventive step when the document is combined with one o r more other such docu¬ ments, such combination being obvious t o a person skilled i n the art "&" document member of the same patent family Date of mailing of the international search report

8 November 2010
Name and mailing address of the ISA/ European Patent Office, P B 5818 Patentlaan 2 N L - 2280 HV RlJSWIjk TeI (+31-70) 340-2040, Fax (+31-70) 340-3016
Form PCT/ISA/210 (second sheet) (April 2005)

12/11/2010
Autho πzed officer

Lechner, Oskar

C(Continuation).

DOCUMENTS

CONSIDERED

T O B E RELEVANT

Category*

Citation o f document,

with indication, where appropriate, o f the relevant passages

Relevant to claim No.

X

MACK MATTHIAS ET AL: "Preferential targeting o f CD4-CCR5 complexes with b i functional inhibitors: a novel approach t o block HIV-I infection." JOURNAL O F IMMUNOLOGY (BALTIMORE, MD. : 1950) 1 DEC 2005 LNKD- PUBMED: 16301668, vol. 175, no. 11, 1 December 2005 (2005-12-01), pages 7586-7593, XP002582205 ISSN: 0022-1767 the whole document SKOV LONE ET AL: "HUMAX-CD4: A FULLY HUMAN MONOCLONAL ANTI-CD4 ANTIBODY FOR THE TREATMENT O F PSORIASIS VULGARIS" ARCHIVES O F DERMATOLOGY, AMERICAN MEDICAL ASSOCIATION, U S LNKDDOI :10 .1001/ARCHDERM .139 .1 1 .1433 , vol. 139, no. 11, 1 November 2003 (2003-11-01), pages 1433-1439, XP009078296 ISSN: 0003-987X the whole document W O 92/05274 A l (GORMAN SCOTT DAVID [GB]; CLARK MICHAEL RONALD [GB]; COBBOLD STEPHEN P ) 2 April 1992 (1992-04-02) claims 10-18 SCHNEIDER-SCHAULIES J : "Cellular receptors for viruses: links t o tropism and pathogenesis." THE JOURNAL O F GENERAL VIROLOGY JUN 2000 LNKD- PUBMED: 10811925, vol. 81, no. Pt 6 , June 2000 (2000-06), pages 1413-1429, XP002582222 ISSN: 0022-1317 the whole document RUDIKOFF S ET AL: "Single amino acid substitution altering antigen-binding specificity" PROCEEDINGS O F THE NATIONAL ACADEMY O F SCIENCES O F THE UNITED STATES (PNAS), NATIONAL ACADEMY O F SCIENCE, U S LNKDDOI :10 .1073/PNAS .79 .6 .1979 , vol. 79, 1 March 1982 (1982-03-01), pages 1979-1983, XP007901436 ISSN: 0027-8424 the whole document

1-5,7-9, 16-18, 20-23,25

X

1-5,7-9, 16-18, 20-23,25

X

1-5,7-9, 16-18, 20-23,25

A

1-5,7-9, 16-18, 20-23,25

A

1-5,7-9, 16-18, 20-23,25

Form PCT/ISA/210 (continuation o f second sheet) (April 2005)

C(Continuatio π ).
Category *

DOCUMENTS CONSIDERED T O B E RELEVANT
Relevant to claim No.

Citation of document, with indication, where appropriate, of the relevant passages

SAERENS D ET AL: "Single-domain antibodies a s building blocks for novel therapeutics" CURRENT OPINION IN PHARMACOLOGY, ELSEVIER SCIENCE PUBLISHERS, N L LNKDDOI: 10. 1016/J. COPH. 2008. 07. 006, vol. 8 , no. 5 , 1 October 2008 (2008-10-01), pages 600-608, XP025609339 ISSN: 1471-4892 [retrieved o n 2008-08-22] the whole document JAIN ET AL: "Engineering antibodies for clinical applications" TRENDS IN BIOTECHNOLOGY, ELSEVIER PUBLICATIONS, CAMBRIDGE, GB, vol. 25, no. 7 , 1 July 2007 (2007-07-01), pages 307-316, XP022116341 ISSN: 0167-7799 the whole document WARK ET AL: "Latest technologies for the enhancement of antibody affinity" ADVANCED DRUG DELIVERY REVIEWS, ELSEVIER BV, AMSTERDAM, N L LNKDDOI: 10. 1016/J. ADDR. 2006. 01. 025, vol. 58, no. 5-6, 7 August 2006 (2006-08-07), pages 657-670, XP005611420 ISSN: 0169-409X the whole document

1-5,7-9, 16-18, 20-23,25

1-5,7-9, 16-18, 20-23,25

1-5,7-9, 16-18, 20-23,25

Form PCT/ISA/210 (continuation of second sheet) (April 2005)

INTERNATIONAL SEARCH REPORT
Box No. Il Observations where certain claims were found unsearchable (Continuation of item 2 of first sheet)

This international search report has not been established in respect of certain claims under Article 17(2)(a) for the following reasons:

1.

Claims Nos.: because they relate to subject matter not required to be searched by this Authority, namely:

Claims Nos.: because they relate to parts of the international application that do not comply with the prescribed requirements to such an extent that no meaningful international search can b e carried out, specifically.

3.

Claims Nos.: because they are dependent claims and are not drafted i n accordance with the second and third sentences of Rule 6.4(a).

Box No. Ill

Observations where unity of invention is lacking (Continuation of item 3 of first sheet)

This International Searching Authority found multiple inventions i n this international application, as follows:

see additi onal sheet

1.

As all required additional search fees were timely paid by the applicant, this international search report covers all searchable claims.

2.

As all searchable claims could be searched without effort justifying a n additional fees, this Authority did not invite payment of additional fees.

3.

As only some of the required additional search fees were timely paid by the applicant, this international search report covers only those claims for which fees were paid, specifically claims Nos.:

4 . _ _ No required additional search fees were timely paid by the applicant. Consequently, this international search report is γ restricted to the invention first mentioned i n the claims; it is covered by claims Nos.:

16-18(completely);
Remark on Protest

1-5, 7-9, 20-23, 25(partially)

The additional search fees were accompanied by the applicant's protest and, where applicable, the payment of a protest fee. The additional search fees were accompanied by the applicant's protest but the applicable protest fee was not paid within the time limit specified in the invitation.
No protest accompanied the payment of additional search fees.

Form PCT/ISA/21 0 (continuation of first sheet (2)) (April 2005)

International Application No. PCT/EP2009

/063411

FURTHER INFORMATION CONTINUED FROM

PCT/ISA/ 210

This International Searching Authority found multiple (groups of) inventions in this international application, a s follows:
1.

claims: 16-18(completely);

1-5, 7-9, 20-23, 25(partially)

Polypeptide comprising one o r more Nanobodies that specifically binds hCD4 consisting of CDRl SeqID 287, CDR2 295 and CDR3 303 and uses thereof.

2 . claims:

19(completely);

1-5, 7-9, 20-23, 25(partially)

Polypeptide comprising one o r more Nanobodies that specifically binds hCD4 consisting of CDRl SeqID 287, CDR2 291 and CDR3 303 and uses thereof.

3.

claims: 1-6, 8-15, 20-25(all partially)

Polypeptide comprising one o r more Nanobodies that specifically binds hCXCR4 and uses thereof characterized by CDRl SeqID142, CDR2 SeqID 174 and CDR3 SeqID 206 6 , 10-15, 24

4 . claims:

1-6, 8-11,

13-15, 20-25(all partially)

Polypeptide comprising one o r more Nanobodies that specifically binds hCXCR4 and uses thereof characterized CDRl SeqID142, CDR2 SeqID 174 and CDR3 SeqID 207

by

5.

claims: 1-6, 8-11, 13-15, 20-25(all partially) Polypeptide comprising one or more Nanobodies that specifically binds hCXCR4 and uses thereof characterized CDRl SeqID142, CDR2 SeqID 175 and CDR3 SeqID 206

by

6 . claims:

1-6, 8-11,

13-15, 20-25(all partially)

Polypeptide comprising one o r more Nanobodies that specifically binds hCXCR4 and uses thereof characterized CDRl SeqID142, CDR2 SeqID 175 and CDR3 SeqID 207

by

7 . claims:

1-6, 8-11,

13-15, 20-25(all partially)

Polypeptide comprising one o r more Nanobodies that specifically binds hCXCR4 and uses thereof characterized CDRl SeqID143, CDR2 SeqID 174 and CDR3 SeqID 206

by

8 . claims:

1-6, 8-11,

13-15, 20-25(all partially)

International Application No. PCT/EP2009

/063411

FURTHER INFORMATION CONTINUED FROM

PCT/ISA/ 210

Polypeptide comprising one o r more Nanobodies that specifically binds hCXCR4 and uses thereof characterized CDRl SeqID143, CDR2 SeqID 174 and CDR3 SeqID 207

by

9 . claims:

1-6, 8-11, 13-15, 20-25(all

partially)

Polypeptide comprising one o r more Nanobodies that specifically binds hCXCR4 and uses thereof characterized CDRl SeqID143, CDR2 SeqID 175 and CDR3 SeqID 206

by

10. claims:

1-6, 8-15, 20-25(all partially)

Polypeptide comprising one o r more Nanobodies that specifically binds hCXCR4 and uses thereof characterized CDRl SeqID143, CDR2 SeqID 175 and CDR3 SeqID 207

by

11. claims:

1-4, 20-23, 25(all partially)

Polypeptide comprising one o r more Nanobodies that specifically binds human alpha2 integrin and uses thereof.

12. claims:

1-4, 20-23, 25(all partially)

Polypeptide comprising one o r more Nanobodies that specifically binds hCCR5 and uses thereof.

13. claims:

1-4, 20-23, 25(all partially)

Polypeptide comprising one o r more Nanobodies that specifically binds human beta3 integrin and uses thereof.

14. claims:

1-4, 20-23, 25(all partially)

Polypeptide comprising one or more Nanobodies that specifically binds human alphaV integrin and uses thereof.

15. claims:

1-4, 20-23, 25(all partially)

Polypeptide comprising one o r more Nanobodies that specifically binds human betal integrin and uses thereof.

16. claims:

1-4, 20-23, 25(all partially)

Polypeptide comprising one o r more Nanobodies that specifically binds human CD81 and uses thereof.

International Application No. PCT/EP2009

/06341 1

FURTHER INFORMATION CONTINUED FROM
17. claims:

POT/ISA/ 2 1 0

1-4, 20-23, 25(all partially)

Polypeptide comprising one o r more Nanobodies that specifically binds human Claudin 1 and uses thereof.

18. claims:

1-4, 20-23, 25(all partially)

Polypeptide comprising one o r more Nanobodies that specifically binds human Claudin 6 and uses thereof.

19. claims:

1-4, 20-23, 25(all partially)

Polypeptide comprising one o r more Nanobodies that specifically binds human Claudin 9 and uses thereof.

Patent document cited in search report

Publication date

Patent family member(s)

Publication date

W O 9205274

Al

02-04-1992

AT AU CA
DE DE DK EP ES GR ID
IE IL

JP NZ US US US ZA

148172 660151 2091769 69124387 69124387 0549581 0549581 2097808 3023252 1044 913257 99499 6502762 239826 6767996 2007212753 7098006 9107407

T
B2 Al Dl T2 T3 Al T3 T3

B
Al

A T A
Bl Al Bl

A

-1997 -1995 -1992 -1997 -1997 -1997 -1993 -1997 -1997 -1996 -1992 -2006 1994 23-12- 1992 27-07- 2004 13-09- -2007 29-08- 2006 24-06- 1992

15-02 15-06 18-03 06-03 15-05 26-05 07-07 16-04 30-07 28-10 25-02 20-08 31-03

Form PCT/ISA/210 (patent family annex) (April 2005)



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