Inhibitor targeting specific complement system, and preparation method and use thereof
An inhibitor targeting specific complement, the inhibitor targeting specific complement is a protein, the protein has a targeted inhibition function on the complement activation, and the amino acid sequence of the protein comprises CRIg extracellular domain and complement inhibiting domain; the amino acid sequence of the protein is consist of the CRIg extracellular domain and the complement inhibiting domain connected directly, or indirectly through a linker which can connect the two domains. The preparation method of the inhibitor targeting specific complement comprises connecting the protein polypeptides of the CRIg extracellular domain and the complement inhibiting domain by gene engineering technology. The protein can be used to prepare a drug targeting inhibition of complement activation. The inhibitor targeting specific complement, its preparation method and applications provided by the present invention, can be used in the treatment and prevention of human diseases caused by complement activation disorders. It has great potential application value and development prospects.
1. An inhibitor protein targeting specific complement comprising an amino acid sequence selected from the group consisting of SEQ ID NO. 2 and SEQ ID NO. 4, and having a targeted inhibition function against complement activation, comprising:
a CRIg extracellular domain amino acid sequence portion; and
a complement inhibiting domain amino acid sequence portion connected to the CRIg extracellular domain amino acid sequence portion, comprising at least a portion of factor H.
2. A nucleic acid which encodes the inhibitor protein targeting specific complement of claim 1 .
3. The nucleic acid according to claim 2 , contained within a vector.
4. The inhibitor according to claim 1 , wherein the CRIg extracellular domain amino acid sequence is directly connected to the complement inhibiting domain amino acid sequence.
5. The inhibitor according to claim 1 , wherein the CRIg extracellular domain amino acid sequence is connected to the complement inhibiting domain amino acid sequence through a linker amino acid sequence.
6. The nucleic acid according to claim 2 , comprising a sequence selected from the group consisting of SEQ ID NO 1 and SEQ ID NO 3.
7. The nucleic acid according to claim 6 , contained within a vector.
8. A method for preparing an inhibitor protein targeting specific complement, comprising:
connecting a nucleic acid sequence encoding a CRIg extracellular domain to a nucleic acid sequence encoding a complement inhibiting domain comprising at least a portion of Factor H, by gene splicing overlap extension PCR, to form a fusion sequence for an inhibitor protein targeting specific complement comprising an amino acid sequence selected from the group consisting of SEQ ID NO. 2 and SEQ ID NO. 4;
inserting the fusion sequence into an eukaryotic expression vector; and
inducing protein expression of the inhibitor protein targeting specific complement through an eukaryotic protein expression vector, the inhibitor protein targeting specific complement comprising the CRIg extracellular domain amino acid sequence portion and the complement inhibiting domain amino acid sequence portion connected to the CRIg extracellular domain amino acid sequence portion.
9. The method according to claim 8 , further comprising purifying the expressed inhibitor protein targeting specific complement.
10. The method according to claim 8 , further comprising the step of administering the inhibitor protein targeting specific complement to a mammal in a pharmaceutically acceptable formulation.
11. A method for treating a condition in a mammal associated with complement activation, comprising administering to the mammal an inhibitor protein targeting specific complement, having a targeted inhibition function against complement activation, comprising an amino acid sequence selected from the group consisting of SEQ ID NO. 2 and SEQ ID NO. 4, comprising:
a CRIg extracellular domain amino acid sequence portion; and
a complement inhibiting domain amino acid sequence portion comprising at least a portion of factor H, connected to the CRIg extracellular domain amino acid sequence portion.
12. The method according to claim 11 , wherein the condition comprises paroxysmal nocturnal hemoglobinuria.
13. The method according to claim 11 , further comprising administering a pharmaceutically acceptable formulation comprising the inhibitor protein targeting specific complement to a mammal.
14. The inhibitor protein targeting specific complement according to claim 1 , wherein the amino acid sequence comprises SEQ ID NO. 2.
15. The inhibitor protein targeting specific complement according to claim 1 , wherein the amino acid sequence comprises SEQ ID NO. 4.
16. The method according to claim 8 , wherein the amino acid sequence comprises SEQ ID NO. 2.
17. The method according to claim 8 , wherein the amino acid sequence comprises SEQ ID NO. 4.
18. The method according to claim 11 , wherein the amino acid sequence comprises SEQ ID NO. 2.
19. The method according to claim 11 , wherein the amino acid sequence comprises SEQ ID NO. 4.