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US 2026/0193632 A1Published application
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.
Claims17
1-19. (canceled)
A method for producing a PH20 variant comprising:
- culturing a host cell comprising a nucleic acid encoding the PH20 variant or an expression vector comprising the nucleic acid,
- wherein the amino acid sequence of the PH20 variant is SEQ ID NO:
- 3 with modifications consisting of:
- (a) an amino acid residue substitution of:
- D65A, E66A, P67A, L68A, N82A, T84N, I102A, D103A, S104A, S104N, I105A, 1105Q, T132A, F133A, Y134A, N166A, N166K, L179A, L179S, L179I, L179F, S180T, S180A, L181A, T182A, T185A, E186A, K187A, V241A, E242A, I243A, K244A, N266A, T267A, Q268A, Q268D, Q268I, Q269A, P271A, V272A, K290A, 1291A, 1291G, 1291L, P292A, P292D, Q311A, V312A, L313A, L313P, L313M, K314A, G340Q, S341H, W342I, W342D, W342H, E343V, E343Y, E343Q, N344F, T345E, T345K, R346M, R346F, R346L, T347Q, T347E, T347V, T347W, T347H, T347S, K348Q, K348F, K348D, E349L, E349W, S350Q, S350I, S350D, Q352E, Q352G, Q352Y, A353E, I354E, I354Q, I354S, I354V, 1354A, 1354N, I354T, I354R, 1354W, K355Q, E356M, E356F, E356I, E356L, E356Q, E356V, Y357W, M358V, M358R, M358Y, M358L, D359K, D359V, D359Y, D359Q, T360Y, T360R, T360L, T361M, T361E, T361H, L362A, N363M, N363E, L441A, S442A, D451A, D451S, T452A, T452D, T452H, T452K, T452G, T452P, T452M, T452F, D453A, D461R, D461A, G462A, V463Y, or V463A relative to SEQ ID NO:
- 3;
- (b) an N-terminus deletion of amino acid residues M1 to T35, M1 to L36, M1 to N37, M1 to F38, M1 to R39, or M1 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 1465 to S490 of SEQ ID NO:
- 3.
8 more elements
A method for producing a PH20 variant comprising:
- culturing a host cell comprising a nucleic acid encoding the PH20 variant or an expression vector comprising the nucleic acid,
- 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 I243 A relative to SEQ ID NO:
- 3;
- (b) an N-terminus deletion of amino acid residues M1 to T35, M1 to L36, M1 to N37, M1 to F38, M1 to R39, or M1 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 1465 to S490 of SEQ ID NO:
- 3.
11 more elements
A method for producing a PH20 variant comprising:
- culturing a host cell comprising a nucleic acid encoding the PH20 variant or an expression vector comprising the nucleic acid,
- 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) N166K, E343V, and T361M relative to SEQ ID NO:
- 3;
- (ii) S350Q and T360R relative to SEQ ID NO:
- 3;
- (iii) N344F, K348Q, and K355Q relative to SEQ ID NO:
- 3;
- (iv) T132S, L181A, E186D, Q268N, I291L, V312A, and T452D relative to SEQ ID NO:
- 3;
- (v) R39K, I105A, T132S, L181M, E186D, Q268A, I291L, V312A, and T452D relative to SEQ ID NO:
- 3;
- (vi) T132A, L181A, E186A, Q268A, I291L, V312A, and T452D relative to SEQ ID NO:
- 3;
- (vii) N344I and K348M relative to SEQ ID NO:
- 3;
- (viii) S341D, W342L, E343S, N344I, T345S, R346S, K348M, K355D, D359E, and T361I relative to SEQ ID NO:
- 3;
- (ix) S341D, W342L, E343S, N344I, T345S, K348M, K355D, D359E, and T361I relative to SEQ ID NO:
- 3;
- (x) T132S, L181A, E186D, Q268N, 1291L, V312A, K348M, and T452D relative to SEQ ID NO:
- 3;
- (xi) T132S, L181A, E186D, Q268N, I291L, V312A, N344I, K348M, and T452D relative to SEQ ID NO:
- 3;
- (xii) 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
- (xiii) 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 M1 to T35, M1 to L36, M1 to N37, M1 to F38, M1 to R39, or M1 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 1465 to S490 of SEQ ID NO:
- 3.
33 more elements
Dependencies are inferred from the claim text.