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JP-7855599-B2 - Alkaline phosphatase polypeptide and method of use thereof

JP7855599B2JP 7855599 B2JP7855599 B2JP 7855599B2JP-7855599-B2

Inventors

  • ヴォエトリ， ウォルター シー．
  • ウー， ユーホン
  • モンテレオーネ， ジョナサン
  • メジェボフスキー， タチアナ
  • ファルコーネ， エリック
  • グオ， ヤン

Assignees

  • アレクシオン ファーマシューティカルズ， インコーポレイテッド

Dates

Publication Date
20260508
Application Date
20220210
Priority Date
20210212

Claims (20)

  1. A pharmaceutical composition comprising a polypeptide having at least 90 % sequence identity with SEQ ID NO: 5 and E108M, N213Q, and N286Q mutations compared to SEQ ID NO: 1, and a pharmaceutically acceptable carrier comprising one or more of phosphate, proline, and sucrose , wherein the polypeptide hydrolyzes inorganic pyrophosphate (PPi) to provide inorganic phosphoric acid (Pi) .
  2. The pharmaceutical composition according to claim 1, wherein the polypeptide has at least 95 %, 97%, or 99% sequence identity with respect to Sequence ID No. 5.
  3. The pharmaceutical composition according to claim 2, wherein the polypeptide comprises or is composed of the sequence of Sequence ID No. 5.
  4. The pharmaceutical composition according to any one of claims 1 to 3 , wherein the composition contains about 1 mM to about 100 mM of phosphate.
  5. The pharmaceutical composition according to claim 4 , wherein the composition comprises about 5 mM to about 20 mM of phosphate.
  6. The pharmaceutical composition according to claim 5 , wherein the composition comprises about 10 mM of phosphate.
  7. The pharmaceutical composition according to any one of claims 1 to 6 , wherein the phosphate is sodium phosphate.
  8. The pharmaceutical composition according to any one of claims 1 to 7 , wherein the composition contains about 1 mM to about 500 mM of proline.
  9. The pharmaceutical composition according to claim 8 , wherein the composition contains about 50 mM to about 200 mM proline.
  10. The pharmaceutical composition according to claim 9 , wherein the composition contains about 140 mM proline.
  11. The pharmaceutical composition according to any one of claims 1 to 10 , wherein the composition contains about 1 mM to about 500 mM sucrose.
  12. The pharmaceutical composition according to claim 11 , wherein the composition contains about 50 mM to about 200 mM sucrose.
  13. The pharmaceutical composition according to claim 12 , wherein the composition contains about 140 mM sucrose.
  14. The pharmaceutical composition according to any one of claims 5 to 13 , wherein the composition comprises about 10 mM phosphate, about 140 mM proline, and about 140 mM sucrose.
  15. The pharmaceutical composition according to any one of claims 1 to 14 , wherein the composition comprises about 0.01% to about 0.5% of sorbitan polyoxyethylene (20) monooleate.
  16. The pharmaceutical composition according to claim 15 , wherein the composition comprises about 0.01% to about 0.1% of sorbitan polyoxyethylene (20) monooleate.
  17. The pharmaceutical composition according to claim 16 , wherein the composition comprises about 0.05% sorbitan polyoxyethylene (20) monooleate.
  18. The pharmaceutical composition according to claim 17 , wherein the composition comprises about 140 mM proline, about 140 mM sucrose, and about 0.05% sorbitan polyoxyethylene (20) monooleate.
  19. The pharmaceutical composition according to any one of claims 1 to 18 , wherein the polypeptide comprises a total sialic acid content (TSAC) of about 1.0 mol/mol to about 6.0 mol/mol.
  20. The pharmaceutical composition according to claim 19 , wherein the TSAC is approximately 3.0 mol/mol to approximately 6.0 mol/mol.

Description

Sequence Listing This application includes a sequence listing submitted electronically in ASCII format, which is incorporated herein by reference in its entirety. The name of the above ASCII copy, created on 4 February 2022, is 50694-094WO2_Sequence_Listing_2_4_22_ST25, and its size is 27,753 bytes. Hypophosphatasia (HPP) is a rare hereditary skeletal disorder with an incidence of 1 in 100,000 live births in the most severe forms of the disease. The disorder is usually caused by loss-of-function mutations in the gene encoding tissue-nonspecific alkaline phosphatase (TNSALP). HPP presents with a very wide range of symptoms and severity, from early tooth loss to near-complete absence of bone mineralization in utero. The symptoms of HPP vary significantly from patient to patient and also significantly with age. Many HPP patients exhibit skeletal changes, short stature, chronic pain, leg pain, gait disturbances, and early non-traumatic tooth loss. Asfotase alfa (STRENSIQ®, Alexion Pharmaceuticals, Inc.), a recombinant enzyme replacement therapy (ERT) containing a soluble fragment of TNSALP, is the first ERT available to HPP patients. Asfotase alfa has demonstrated groundbreaking effects on the most severe forms of HPP, as evidenced by improvements in bone mineralization and density, as well as respiratory and motor function, cognitive development, and muscle strength (White et al., New Engl. J.Med. 366:904-913, 2012). White et al. , New Engl. J. Med. 366:904-913, 2012 The first embodiment is characterized by a pharmaceutical composition comprising an alkaline phosphatase polypeptide having at least 80% (e.g., at least 85%, 90%, 95%, 97%, 99%, or 100%) sequence identity with SEQ ID NO: 5, and a pharmaceutically acceptable carrier. The polypeptide may contain at least one mutation selected from E108M, N213Q, and N286Q compared to the amino acid sequence of SEQ ID NO: 1 (e.g., the polypeptide may contain two or all three of these mutations). For example, the polypeptide has at least 80% (e.g., at least 85%, 90%, 95%, 97%, 99%, or 100%) sequence identity with the amino acid sequence of SEQ ID NO: 5 and contains at least one, two, or all three of the mutations selected from E108M, N213Q, and N286Q. The pharmaceutically acceptable carrier may contain one or more of phosphates, proline, and sucrose. For example, a polypeptide may contain or be composed of the amino acid sequence of SEQ ID NO: 5. In some embodiments, the alkaline phosphatase moiety of the polypeptide has at least 80% (e.g., at least 85%, 90%, 95%, 97%, 99%, or 100%) sequence identity with SEQ ID NO: 3. This polypeptide may be further linked to an Fc region (e.g., an IgG1, IgG2, IgG3, or IgG4Fc region) and/or a polyaspartic acid region. In some embodiments, the polypeptide includes an IgG2/4Fc region and has at least 80% (e.g., at least 85%, 90%, 95%, 97%, 99%, or 100%) identity with SEQ ID NO: 4. The polyaspartic acid may include, for example, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 aspartic acid residues. In some embodiments, polyaspartic acid contains 10 aspartic acid residues (D10). The composition may be prepared to contain an alkaline phosphatase polypeptide in doses ranging from about 0.1 mg/mL to about 200 mg/mL (e.g., about 1, 10, 20, 25, 50, 75, 100, 125, 150, 175, or 200 mg/mL). The composition may be prepared in volumes ranging from about 0.1 mL to about 50 mL (e.g., about 0.1 to about 10 mL, e.g., about 0.1 mL, 0.2 mL, 0.3 mL, 0.4 mL, 0.5 mL, 0.6 mL, 0.7 mL, 0.8 mL, 0.9 mL, or 1.0 mL, e.g., about 1 mL to about 10 mL, e.g., about 2 mL, 3 mL, 4 mL, 5 mL, 6 mL, 7 mL, 8 mL, 9 mL, or 10 mL). In some embodiments, the composition is prepared in about 1 mL. For example, the composition may contain at least 80% (e.g., at least 85%, 90%, 95%, 97%, or 99%) sequence identity with respect to SEQ ID NO: 5, or 100 mg/mL of alkaline phosphatase polypeptide having that sequence (e.g., the polypeptide contains at least one, two, or all three mutations selected from E108M, N213Q, and N286Q of SEQ ID NO: 5), and a pharmaceutically acceptable carrier. The composition may contain a phosphate (e.g., sodium phosphate) at a concentration of, for example, about 1 mM to about 100 mM, or about 5 mM to about 20 mM, for example, about 2 mM, 3 mM, 4 mM, 5 mM, 6 mM, 7 mM, 8 mM, 9 mM, 10 mM, 20 mM, 30 mM, 40 mM, 50 mM, 60 mM, 70 mM, 80 mM, 90 mM, or 100 mM, for example, about 10 mM. The composition may further contain proline and/or sucrose. The composition may further contain proline. The composition may further contain sucrose. For example, the composition contains proline at concentrations of approximately 1 mM to 500 mM, approximately 70 mM to 280 mM, approximately 50 mM to 200 mM, approximately 2 mM, 3 mM, 4 mM, 5 mM, 6 mM, 7 mM, 8 mM, 9 mM, 10 mM, 20 mM, 30 mM, 40 mM, 50 mM, 60 mM, 70 mM, 80 mM, and 90 mM. 100 mM, 110 mM, 120 mM, 130 mM, 140 mM, 150 mM, 160 mM, 170 mM, 180 mM, 190 mM, 200 mM, 300 mM, 400 mM, or 500 mM, for example, about 14