Mutated Netrin-4, Fragments Thereof And Their Use As Medicines

• US 20100183588A1
• Filed: 11/22/2005
• Published: 07/22/2010
• Est. Priority Date: 11/22/2004
• Status: Active Grant

• Current Assignee: Centre National De La Recherche Scientifique, IVS Institut Des Vaisseaux Et Du Sang
• Original Assignee: Centre National De La Recherche Scientifique, IVS Institut Des Vaisseaux Et Du Sang
• Inventors: Plouet, Jean, Alemany, Monica
• Granted Patent: US 7,999,072 B2
• Time in Patent Office: Days
• US Class Current: 424/131.100
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 17/06 Antipsoriatics
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 19/08 for bone diseases, e.g. rachitism, Paget's disease
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: A61P 27/06 Antiglaucoma agents or miotics
• CPC Class Codes: A61P 27/10 for accommodation disorders, e.g. myopia
• 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 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 3/04 Anorexiants; Antiobesity agents
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: A61P 37/02 Immunomodulators
• 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 9/10 : for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: A61P 9/14 : Vasoprotectives; Antihaemorrhoidals; Drugs for varicose therapy; Capillary stabilisers
• CPC Class Codes: C07K 14/475 : Growth factors; Growth regulators
• CPC Class Codes: C07K 16/22 : against growth factors ; against growth regulators

Abstract

Described is use of a netrin selected among netrin-4, mutated netrin-4, netrin-1, netrin G1 or netrin-3, or one of their fragments or a nucleotide sequence encoding one of the netrins or one of the fragments, or an anti-idiotypic antibody of one of the netrins or of one of the fragments, or of a Fab fragment of the anti-idiotypic antibodies, for preparing a medicine for preventing or treating tumoral or non-tumoral pathologies.

Claims

1. Protein comprising or being constituted by one of the following sequences:

the sequence SEQ ID NO;

522 or SEQ ID NO;

524, or

a fragment of said sequence SEQ ID NO;

522 or SEQ ID NO;

524, providing that this fragment exhibits anti-angiogenic activity and/or pericyte activation activity, said fragment comprising 40 to 450 amino acids, and preferably 40 to 260 amino acids and preferably 40 to 230 amino acids, and being represented by one of the sequences SEQ ID NO;

2q, q varying from 187 to 248, or by the sequence SEQ ID NO;

526 or by the sequence SEQ ID NO;

528,

any sequence derived from one of the abovementioned sequences, by substitution, suppression or addition of one or more amino acids, providing that this derived sequence exhibits anti-angiogenic activity and/or pericyte activation activity, or

any sequence homologous to one of the abovementioned sequences, preferably having at least 50% identity with the region comprised between the amino acids at position 261 and 515 of SEQ ID NO;

522, providing that this homologous sequence exhibits anti-angiogenic activity and/or pericyte activation activity.

2. Protein according to claim 1 comprising or being constituted by one of the following sequences:

the sequence SEQ ID NO;

522 or SEQ ID NO;

524, or

a fragment of said sequence SEQ ID NO;

522 or SEQ ID NO;

524, providing that this fragment exhibits anti-angiogenic activity, said fragment comprising 40 to 450 amino acids, and preferably 40 to 260 amino acids and preferably 40 to 230 amino acids, and being represented by one of the sequences SEQ ID NO;

2q, q varying from 187 to 248, or by the sequence SEQ ID NO;

526 or by the sequence SEQ ID NO;

528,

any sequence derived from one of the abovementioned sequences, by substitution, suppression or addition of one or more amino acids, providing that this derived sequence exhibits anti-angiogenic activity, or

any sequence homologous to one of the abovementioned sequences, preferably having at least 50% identity with the region comprised between the amino acids at position 261 and 515 of SEQ ID NO;

522, providing that this homologous sequence exhibits anti-angiogenic activity.

3. Protein according to claim 1, comprising or being constituted by one of the following sequences:

the sequence SEQ ID NO;

522 or SEQ ID NO;

524, or

a fragment of said sequence SEQ ID NO;

522 or SEQ ID NO;

524, providing that this fragment exhibits pericyte activation activity, said fragment comprising 40 to 450 amino acids, and preferably 40 to 230 amino acids, and preferably 40 to 260 amino acids and being represented by one of the sequences SEQ ID NO;

2q, q varying from 187 to 248, or by the sequence SEQ ID NO;

526 or by the sequence SEQ ID NO;

528,

any sequence derived from one of the abovementioned sequences, by substitution, suppression or addition of one or more amino acids, providing that this derived sequence exhibits pericyte activation activity, or

any sequence homologous to one of the abovementioned sequences, preferably having at least 50% identity with the region comprised between the amino acids at position 261 and 515 of SEQ ID NO;

522, providing that this homologous sequence exhibits pericyte activation activity.

4. Nucleotide sequence coding for a protein according to claim 1.

5. Nucleotide sequence according to claim 4, comprising or being constituted by:

the nucleotide sequence SEQ ID NO;

521 coding for SEQ ID NO;

522, or the nucleotide sequence SEQ ID NO;

523 coding for SEQ ID NO;

524,

a fragment of one of these nucleotide sequences, and represented by one of the sequences SEQ ID NO;

2q−

1, q varying from 187 to 248, or by the sequence SEQ ID NO;

525 or by the sequence SEQ ID NO;

527,

or any nucleotide sequence derived, by degeneration of the genetic code, from one of the abovementioned nucleotide sequences, and coding for a protein represented by one of the sequences SEQ ID NO;

2q, q varying from 187 to 248, or by the sequence SEQ ID NO;

526 or the sequence SEQ ID NO;

528,

or any nucleotide sequence derived, by substitution, suppression or addition of one or more nucleotides, from one of the abovementioned nucleotide sequences coding for a protein derived from SEQ ID NO;

2q, q varying from 187 to 248, or for the sequence SEQ ID NO;

526 or for the sequence SEQ ID NO;

528, as defined above,

or any nucleotide sequence homologous to one of the abovementioned nucleotide sequences, preferably having at least 50% identity with one of the sequences SEQ ID NO;

2q−

1 coding for a protein homologous to SEQ ID NO;

2q, or with the sequence SEQ ID NO;

525 coding for the sequence SEQ ID NO;

526 or with sequence SEQ ID NO;

527 coding for the sequence SEQ ID NO;

528, as defined above,

or any nucleotide sequence complementary to the abovementioned sequences,

or any nucleotide sequence capable of hybridizing under stringent conditions with the sequence complementary to one of the abovementioned sequences.

6. Recombinant vector, chosen among plasmid, cosmid, phage or virus DNA, containing a nucleotide sequence as defined in claim 4, and containing the elements necessary for the expression in a host cell of the polypeptides encoded by the nucleic acids abovementioned, inserted into said vector.

7. Host cell chosen from bacteria, viruses, yeasts, fungi, plants or mammal cells, said host cell being transformed, using a recombinant vector as defined in claim 6.

8. Antibody being directed specifically against a protein as defined in claim 1.

9. Anti-idiotypic antibody of a protein as defined in claim 1.

10. Fab fragment of anti-idiotypic antibodies as defined in claim 9.

11. Pharmaceutical composition, comprising as active ingredient:

a protein as defined in claim 1.

12. A method for the prevention or treatment of pathologies requiring the inhibition of endothelial proliferation and/or migration, within the context of the following pathologies:

cancers and leukemias, choroidal neovascularization complicating myopia, neovascularization of the cornea like graft rejection, glaucoma, diabetic retinopathies or retinopathies of premature infants, rheumatoid polyarthritis, psoriatic polyarthritis, angiomas, angiosarcomas, Castelman'"'"'s disease and Kaposi'"'"'s sarcoma, or within the context of the treatment of obesity or retinal neovascularisation, said method comprising the administration to a patient in need thereof,

of a protein as defined in claim 1.

13. A method for the prevention or treatment of pathologies requiring the stimulation of endothelial proliferation and/or migration, within the context of the following pathologies:

ischemic pathologies such as arteritis of the lower limbs, myocardial infarction, cerebral vascular accidents, sclerodermia or Raynaud'"'"'s disease, said method comprising the administration to a patient in need thereof,

of an antibody as defined in claim 8.

14. A method for the prevention or treatment of the non-tumoral pathologies linked to, or caused by, a rarefaction of the pericytes or the smooth muscle cells, said method comprising the administration to a patient in need thereof,

of a protein as defined in claim 1.

15. A method according to claim 14, wherein the non-tumoral pathologies linked to, or caused by, a rarefaction of the pericytes or the smooth muscle cells, and requiring activation of the proliferation or migration of the pericytes or the smooth muscle cells, are chosen from:

age-related macular degeneration, choroidal neovascularization complicating myopia,

diabetic retinopathies or retinopathies of premature infants,

neovascular glaucoma,

neovascularization of the cornea in particular graft rejection

rheumatoid polyarthritis,

psoriasis, in particular psoriatic polyarthritis,

angiomas,

atherosclerosis,

obesity,

intestinal malformations,

Crohn'"'"'s disease,

vascular, sub-cortical vascular dementia Alzheimer'"'"'s disease,

degenerative bone pathologies and fractures, and

aneurysms and vascular dissections.

23. A method for the prevention or treatment of tumoral or non-tumoral pathologies as defined in claim 12, said method comprising the administration to a patient in need thereof,

of mutated netrin 4 represented by SEQ ID NO:

522 or SEQ ID NO;

524, or

of a fragment of said sequence SEQ ID NO;

522 or SEQ ID NO;

524, represented by one of the sequences SEQ ID NO;

2q, q varying from 187 to 248, or by the sequence SEQ ID NO;

526 or the sequence SEQ ID NO;

528,

in combination with an anti-angiogenic agent chosen from;

AVASTIN (bevacizumab), MACUGEN (pegaptanib), and LUCENTIS (ranibizumab), or any other anti-VEGF agent.

24. Combination product comprising:

mutated netrin 4 represented by SEQ ID NO;

522 or SEQ ID NO;

524, or

a fragment of said sequence SEQ ID NO;

522 or SEQ ID NO;

524, represented by one of the sequences SEQ ID NO;

2q, q varying from 187 to 248, or by the sequence SEQ ID NO;

526 or the sequence SEQ ID NO;

528,

in combination with an anti-angiogenic agent AVASTIN (bevacizumab), MACUGEN (pegaptanib) and LUCENTIS (ranibizumab), or any other anti-VEGF agent for simultaneous or separate use or spread over time, intended for the treatment or prevention of tumoral or non-tumoral pathologies as defined in claim 12.

16. A method for the treatment of cancer, said method comprising the administration to a patient in need thereof,

of mutated netrin 4 represented by SEQ ID NO:

522 or SEQ ID NO;

524, or

of a fragment of said sequence SEQ ID NO;

522 or SEQ ID NO;

524, represented by one of the sequences SEQ ID NO;

2q, q varying from 187 to 248, or by the sequence SEQ ID NO;

526 or by the sequence SEQ ID NO;

528,

in combination with a chemotherapy agent.

17. Protein, comprising or being constituted by one of the following sequences:

a sequence SEQ ID NO;

2i, i varying from 64 to 131, or

any sequence derived from one of the abovementioned sequences, by substitution, suppression or addition of one or more amino acids, providing that this derived sequence exhibits anti-angiogenic activity, or

any sequence homologous to one of the abovementioned sequences, preferably having at least 50% identity with the region comprised between the amino acids at position 284 and 487 of SEQ ID NO;

502, providing that this homologous sequence exhibits anti-angiogenic activity, providing that said protein is different from the sequence SEQ ID NO;

502 or SEQ ID NO;

504.

18. Nucleotide sequence coding for a protein according to claim 17.

19. Nucleotide sequence according to claim 18, comprising or being constituted by:

the nucleotide sequence SEQ ID NO;

2i−

1, i varying from 64 to 131, or

any nucleotide sequence derived, by degeneration of the genetic code, from one of the abovementioned nucleotide sequences, and coding for a protein represented by one of the sequences SEQ ID NO;

2i, i varying from 64 to 131, or

any nucleotide sequence derived, by substitution, suppression or addition of one or more nucleotides, from one of the abovementioned nucleotide sequences coding for a protein derived from one of the sequences SEQ ID NO;

2i, i varying from 64 to 131, as defined above, or

any nucleotide sequence homologous to one of the abovementioned nucleotide sequences, preferably having at least 50% identity with one of the sequences SEQ ID NO;

2i−

1 coding for a protein homologous to SEQ ID NO;

2i, i varying from 64 to 131, as defined above, or

any nucleotide sequence complementary to the abovementioned sequences, or

any nucleotide sequence capable of hybridizing under stringent conditions with the sequence complementary to one of the abovementioned sequences,

providing that said nucleotide sequence is different from the sequence

SEQ ID NO;

501 or SEQ ID NO;

503.

20. Pharmaceutical composition, comprising as active ingredient:

a protein as defined in claim 17, or

an antibody, characterized in that it is being directed specifically against a protein as defined in claim 17, or

an anti-idiotypic antibody of a protein as defined in claim 17, or

a Fab fragment of anti-idiotypic antibodies of a protein as defined in claim 17.

21. A method for the prevention or treatment of pathologies requiring the inhibition of endothelial proliferation and/or migration, within the context of the following pathologies:

cancers and leukemias, choroidal neovascularization complicating myopia, neovascularization of the cornea like graft rejection, glaucoma, diabetic retinopathies or retinopathies of premature infants, rheumatoid polyarthritis, psoriatic polyarthritis, angiomas, angiosarcomas, Castelman'"'"'s disease and Kaposi'"'"'s sarcoma, or within the context of the treatment of obesity or retinal neovascularisation, said method comprising the administration to a patient in need thereof,

of a protein as defined in claim 17, or

of an anti-idiotypic antibody of a protein as defined in claim 17.

22. A method for the prevention or treatment of pathologies requiring the stimulation of endothelial proliferation and/or migration, within the context of the following pathologies:

ischemic pathologies such as arteritis of the lower limbs, myocardial infarction, cerebral vascular accidents, sclerodermia or Raynaud'"'"'s disease, said method comprising the administration to a patient in need thereof,

of an antibody, being directed specifically against a protein as defined in claim 17, or

of a Fab fragment of anti-idiotypic antibodies of a protein as defined in claim 17.