Novel hyaluronidase variants with improved stability and pharmaceutical composition comprising the same
AssigneeAlteogen, Inc.
InventorsSoon Jae PARK, Hye-Shin CHUNG, Seung Joo LEE, Kyuwan KIM, Hyung-Nam SONG, Sun-Ah YOU, Chang Woo LEE
Life of this patent
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Abstract
Disclosed are novel PH20 variants or fragments thereof with improved thermal stability and enzymatic activity of human hyaluronidase, which is an enzyme that hydrolyzes hyaluronic acid, and more particularly novel PH20 variants or fragments thereof including one or more amino acid residue substitutions in the variant having the amino acid sequence of SEQ ID NO: 3, wherein one or more amino acid residues at the N-terminus and/or the C-terminus are optionally further deleted.
Claims25
1-19. (canceled)
A PH20 variant, wherein the amino acid sequence of the PH20 variant is SEQ ID NO:
- 3 with modifications consisting of:
- (a) an amino acid residue substitution relative to SEQ ID NO:
- 3, selected from the following (i)-(iv):
- (i) D359Q, T361M, I354T, E343Q, N166K, Q352E, A353E, K355Q, D359K, T360Y, N344F, I354A, I354N, N363E, S350Q, R346M, Q352Y, E356Q, E356F, S350D, T360L, W342H, K348Q, N363M, or M358Y;
- (ii) I354V, D359V, Q352G, T361H, E356M, I354E, I354Q, D359Y, I354S, T361E, or I354R;
- (iii) Q311A or D451A;
- or
- (iv) L313M, T452D, S180T, V463Y, I105Q, T452H, T452A, T452K, L179A, Q268I, T452G, V463A, P292D, T452P, D451S, D103A, I291G, T452F, L179F, L313P, L179S, L179I, D461R, or G462A;
- (b) an N-terminus deletion of amino acid residues Ml to T35, Ml to L36, Ml to N37, Ml to F38, Ml to R39, or Ml to A40 of SEQ ID NO:
- 3;
- and
- (c) a C-terminus deletion, wherein the C-terminus of the PH20 variant ends with an amino acid residue selected from any one of amino acid residues I465 to S490 of SEQ ID NO:
- 3.
A PH20 variant, wherein the amino acid sequence of the PH20 variant is SEQ ID NO:
- 3 with modifications consisting of:
- (a) an amino acid residue substitution selected from the following (i) or (ii):
- (i) E356L, E356I, T84N, K348F, E356V, or T347H relative to SEQ ID NO:
- 3;
- or
- (ii) T267A, D461A, N166A, V312A, I291A, Q268A, I105A, T132A, E186A, T182A, L181A, V272A, S104A, T185A, S180A, D65A, L313A, P292A, P67A, P271A, D453A, S442A, K187A, F133A, E66A, L441A, K290A, N82A, K244A, K314A, or I243A relative to SEQ ID NO:
- 3;
- (b) an N-terminus deletion of amino acid residues Ml to T35, Ml to L36, Ml to N37, Ml to F38, Ml to R39, or Ml to A40 of SEQ ID NO:
- 3;
- and
- (c) a C-terminus deletion, wherein the C-terminus of the PH20 variant ends with an amino acid residue selected from any one of amino acid residues I465 to S490 of SEQ ID NO:
- 3.
A PH20 variant, wherein the amino acid sequence of the PH20 variant is SEQ ID NO:
- 3 with modifications consisting of:
- (a) amino acid residue substitutions in one of the following groups:
- (i) S350Q and T360R relative to SEQ ID NO:
- 3;
- (ii) N344F, K348Q, and K355Q relative to SEQ ID NO:
- 3;
- (iii) T132S, L181A, E186D, Q268N, 1291L, V312A, and T452D relative to SEQ ID NO:
- 3;
- (iv) R39K, I105A, T132S, L181M, E186D, Q268A, I291L, V312A, and T452D relative to SEQ ID NO:
- 3;
- (v) T132A, L181A, E186A, Q268A, I291L, V312A, and T452D relative to SEQ ID NO:
- 3;
- (vi) N344I and K348M relative to SEQ ID NO:
- 3;
- (vii) S341D, W342L, E343S, N344I, T345S, R346S, K348M, K355D, D359E, and T361I relative to SEQ ID NO:
- 3;
- (viii) S341D, W342L, E343S, N344I, T345S, K348M, K355D, D359E, and T361I relative to SEQ ID NO:
- 3;
- (ix) T132S, L181A, E186D, Q268N, I291L, V312A, K348M, and T452D relative to SEQ ID NO:
- 3;
- (x) T132S, L181A, E186D, Q268N, I291L, V312A, N344I, K348M, and T452D relative to SEQ ID NO:
- 3;
- (xi) T132A, L181A, E186A, Q268A, I291L, V312A, S341D, W342L, E343S, N344I, T345S, R346S, K348M, K355D, D359E, T361I, and T452D relative to SEQ ID NO:
- 3;
- or
- (xii) T132A, L181A, E186A, Q268A, I291L, V312A, S341D, W342L, E343S, N344I, T345S, K348M, K355D, D359E, T361I, and T452D relative to SEQ ID NO:
- 3;
- (b) an N-terminus deletion of amino acid residues Ml to T35, Ml to L36, Ml to N37, Ml to F38, Ml to R39, or Ml to A40 of SEQ ID NO:
- 3;
- and
- (c) a C-terminus deletion, wherein the C-terminus of the PH20 variant ends with an amino acid residue selected from any one of amino acid residues I465 to S490 of SEQ ID NO:
- 3.
Dependencies are inferred from the claim text.