JP2005538931A5

JP2005538931A5 -

Info

Publication number: JP2005538931A5
Application number: JP2003566050A
Authority: JP
Filing date: 2003-02-07
Publication date: 2006-03-30

Claims

a) a molecular weight of about 65-70 kDa, more preferably 66-69 kDa,
b) pI is greater than 10 or less than 3,
c) when bound to the rhamnose portion of solamargin, the dissociation constant is about 1.5 × 10 ⁻⁶ ;
d) configured to bind to a rhamnose affinity column prepared according to Example 1 under the conditions described in Example 1;
e) configured to elute from the column described in Example 1 using 100 mM rhamnose solution;
f) An isolated RBP that is insoluble in aqueous solution, having at least one characteristic of being dissolved in a highly denaturing buffer containing greater than about 2% surfactant .

a) a molecular weight of about 65-70 kDa, more preferably 66-69 kDa,
b) when bound to the rhamnose portion of solamargin, the dissociation constant is about 1.5 × 10 ⁻⁶ ;
c) configured to bind to a rhamnose affinity column prepared according to Example 1 under the conditions described in Example 1;
d) configured to elute from the column described in Example 1 using 100 mM rhamnose solution;
e) An isolated RBP that is insoluble in aqueous solution with at least one characteristic of dissolving in a highly denaturing buffer containing greater than about 2% surfactant .

The isolated RBP of claim 1 or 2, wherein the molecular weight is about 67 kDa.

The method for identifying a compound that binds to RBP according to any one of claims 1 to 3, comprising a step of bringing a candidate compound into contact with RBP and a step of evaluating binding.

5. The method of claim 4, wherein the compound is an agonist selected from the group consisting of peptides, antibodies and fragments thereof, and peptidomimetics.

5. The method of claim 4, wherein the compound is an antagonist selected from the group consisting of peptides, antibodies and fragments thereof, and peptidomimetics.

4. RBP (or a part thereof ) according to any one of claims 1 to 3, comprising contacting a candidate compound with rhamnose and RBP and assessing whether the compound competes with RBP for rhamnose. And a method for identifying a compound that binds to rhamnose.

The candidate compound is
(I) members of soluble and random peptide libraries or molecular libraries derived from combinatorial chemistry made from D-type and / or L-type amino acids; (ii) phosphopeptides (random or partially modified, present Including, but not limited to, members of a directed phosphopeptide library), (iii) antibodies (polyclonal antibodies, monoclonal antibodies, humanized antibodies, anti-idiotype antibodies, chimeric antibodies, or single chain antibodies; FAb, F ( ab ') ₂ and FAb expression library fragments and epitope binding fragments thereof, including but not limited to, (iv) selected from the group consisting of small organic or inorganic small molecules The method of any one of these.

Use of a compound identified using the method according to any one of claims 4 to 8 for determining the biological function of RBP or treating cancer.

Use of a compound identified using the method according to any one of claims 4 to 6 for isolating RBP homologues.

Identified using the method of claim 7 for modifying the activity of wild-type and / or mutant RBP, elucidating the biological function of RBP, or screening for compounds that disrupt normal RBP interactions Use of compounds.

(I) detecting the level of RBP according to any one of claims 1 to 3 in the sample, and (ii) comparing the level of RBP in the non-cancer-derived sample. Cancer detection method.

(I) detecting the level of RBP according to any one of claims 1 to 3 in a sample derived from a patient, and (ii) comparing the level of RBP in a sample derived from non-cancer. How to diagnose cancer in a patient.

(I) a step of detecting the level and / or distribution of RBP according to claim 1 of a patient, a sign of cancer by analyzing the distribution and / or the level of (ii) the RBP A method of diagnosing cancer in a patient, comprising identifying a difference.

A biological fluid incubated in a cell line in the presence of a detectably labeled antibody capable of identifying the RBP of any one of claims 1 to 3 or a conservative variant, peptide or fragment thereof, An immunoassay comprising incubating a sample such as a tissue extract, freshly harvested cells, or cell lysate and detecting the bound antibody.

Use of an RBP agonist for delivering a drug to a cell having a higher load according to any one of claims 1-3, such as a cancer cell.

4. An agonist of RBP according to any one of claims 1 to 3 linked to an agent configured to prevent cell growth or division or cause cell death.

18. The agent of claim 17, wherein the agent is selected from the group consisting of doxorubicin, daunorubicin, vincristine, vinblastine, vindesine, methotrexate, cytarabine, etopside, cisplatin, carboplatin, 5-fluorouracil, bleomycin, epirubicin, cyproterone, and irinotecan. RBP agonist-drug conjugate.

Use of an RBP agonist according to claim 17 or 18 for the treatment of cancer in a patient.

19. Use of an RBP agonist according to claim 17 or 18 for the treatment of BEC (R) overdose.

21. Use according to claim 19 or 20, wherein the agonist is an RBP antibody, rhamnose, or other RBP ligand.

A pharmaceutical or veterinary composition comprising an agonist of RBP according to any one of claims 1 to 3 and a pharmaceutically acceptable carrier.

24. The pharmaceutical composition of claim 22, further comprising an agent configured to cause cell death, such as a glycoside.

24. A pharmaceutical composition according to claim 22 or 23 adapted for topical administration to a patient.

25. A pharmaceutical composition according to claim 24 in the form of a solution, suspension, emulsion, cream, gel, ointment, powder, poultice, salve, aerosol, or transdermal patch. .

One or more epitopes of RBP according to claim 1, wherein the RBP epitopes of conserved variants or antibodies specifically recognizing the peptide fragment of the RBP,.

Polyclonal antibodies, monoclonal antibodies, humanized antibodies, chimeric antibodies, single chain antibodies, Fab fragments, F (ab ′) ₂ fragments, fragments produced by Fab expression libraries, anti-idiotype (anti-Id) antibodies, and the above 27. The antibody of claim 26, comprising any epitope binding fragment.

28. Use of an antibody according to claim 26 or 27 for detecting RBP in a biological sample.