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KR-102960889-B1 - Vaccine adjuvants and formulations

KR102960889B1KR 102960889 B1KR102960889 B1KR 102960889B1KR-102960889-B1

Abstract

A composition comprising an antigen, a carbohydrate, and a metabolic oil, a method of administering such composition to a subject, a method of preparing such compound, and related compositions, methods, and uses.

Inventors

  • 투오히, 빈센트, 케빈
  • 존슨, 저스틴, 엠.

Assignees

  • 더 클리브랜드 클리닉 파운데이션

Dates

Publication Date
20260507
Application Date
20200213
Priority Date
20190215

Claims (20)

  1. (a) Tumor-associated antigen; (b) Zymosan; (c) mineral oil; and (d) A composition comprising mandib monooleate.
  2. A composition according to claim 1, capable of inducing an antigen-specific T cell immune response including both type 1 and type 17 proinflammatory T cell responses when administered to a subject.
  3. A composition according to claim 1, wherein the composition is a water-in-oil emulsion.
  4. A composition according to claim 1, wherein the tumor-associated antigen is a retired auto-antigen.
  5. A composition according to claim 4, wherein the tumor-associated antigen comprises an α-lactalbumin polypeptide comprising eight or more consecutive amino acids of SEQ ID NO. 5.
  6. A composition according to claim 1, wherein the ratio of antigen to zymosan in the composition is 10:1 to 1:10 (w/w).
  7. A composition according to claim 5, wherein the α-lactalbumin polypeptide comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO. 5.
  8. In claim 7, the α-lactalbumin polypeptide is a composition comprising the amino acid sequence of SEQ ID NO. 5.
  9. In claim 5, the composition is a water-in-oil emulsion.
  10. In claim 8, the composition is a water-in-oil emulsion.
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Description

Vaccine adjuvants and formulations Related applications This application claims priority to U.S. Provisional Patent Application No. 62/806,422 filed on February 15, 2019, the entire contents of which are incorporated by reference for all purposes. Sequence list This specification refers to the sequence list (electronicly submitted on February 13, 2020, as a .txt file named "CCI_005_Seq_Listing.txt"). This .txt file was created on February 13, 2020, and is 6kb in size. The full contents of the sequence list are incorporated herein by reference. background Vaccines generally contain at least two major components: an immunogen that serves as a target for the acquired immune response, and an adjuvant that enhances the acquired immune response. Complete Freund's Adjuvant (CFA) is a suspension of dead mycobacteria in a liquid prepared from non-metabolic oils. CFA is widely regarded as the "gold standard" compared to all other adjuvants due to its proven efficacy in inducing acquired immunity over more than 70 years. However, CFA cannot be used as an adjuvant in human vaccination due to toxic effects primarily associated with the induction of unresolved granulomas and abscesses at the injection site. Therefore, an adjuvant suitable for human vaccination that can promote the induction of a strong immune response is required. Breast cancer is the second most common cause of cancer-related death in women. Breast cancer has several different genetic subtypes, and treatment is generally directed toward specific subtypes. For example, hormone therapies and drugs targeting the estrogen receptor (ER) are designed to treat ER-positive cancers. Triple-negative breast cancer (TNBC) is the most aggressive and deadliest form of breast cancer and is notoriously difficult to treat. Since TNBC cancer cells are negative for the estrogen receptor (ER), progesterone (PR), and HER2, drugs designed to target these receptors are ineffective against TNBC. Effective breast cancer treatments are needed for hard-to-treat breast cancer subtypes, including TNBC. Figure 1 shows the induction of pro-inflammatory T cell immunity by vaccination with an α-lactalbumin/complete Freund adjuvant (CFA) emulsion. The mean splenocyte frequency of IFNγ (Type 1) and IL-17 (Type 17)-producing pro-inflammatory T cells was determined 4 weeks after vaccination of 6 to 8-week-old BALB/c female mice (n=3) with 200 μg of Mycobacterium tuberculosis strain H37RA containing complete Freund adjuvant (CFA) and 100 μg of recombinant mouse α-lactalbumin. Error bars represent ±SD. Figure 2 shows a comparison of CFA and various adjuvants in inducing type 1/type 17 T cell immunity. Splenocyte frequencies of type 1 and type 17 pro-inflammatory T cells were measured after 4 weeks of immunizing 6 to 8-week-old BALB/cJ female mice with 200 μL of emulsion containing various adjuvants including 100 μg of recombinant mouse α-lactalbumin and a standard dose of CFA. Data are expressed as the mean α-lactalbumin-specific type 1 and type 17 spot-forming units (SFU) obtained using each test adjuvant divided by the SFU obtained on the same day using CFA as the "gold standard" adjuvant. The horizontal black dashed line represents the same frequency as that obtained using CFA as an adjuvant (SFU induced using CFA = 1). Some immunizations were performed twice at a 2-week interval and are indicated as x2. Figures 3a to 3h show the growth of 4T1 mouse mammary tumors using different α-lactalbumin/adjuvant combinations. Figure 3a shows female BALB/cJ mice subcutaneously vaccinated at 6 to 8 weeks of age with 200 μL of an emulsion containing 100 μg recombinant mouse α-lactalbumin and 200 μg CFA. Control mice were vaccinated with 200 μg CFA alone. Two weeks after vaccination, 2 x 10⁴ 4T1 mouse mammary tumor cells were subcutaneously inoculated into the mice, and tumor growth was measured every other day using a vernier caliper. Subsequently, the same protocol was used to determine tumor growth using the manufacturer-recommended doses for various other adjuvants shown in Fig. 2, including GPI-0100 x2 (Fig. 3b), Sigma lipid A (Fig. 3c), AS02B lipid A (Fig. 3d), CpG DNA x2 (Fig. 3e), CpG DNA + α-Gal-Cer x2 (Fig. 3f), β-glucan peptide in IFA (Fig. 3g), and zymosan in IFA (Fig. 3h). Some vaccinations were performed twice at a 2-week interval and are indicated as x2. An asterisk indicates a significant difference (P<0.05) between the test and control vaccinated mice. Figure 4 shows the induction of type 1/type 17 pro-inflammatory T cells using Zymosan/IFA versus Zymosan/MONTANIDE ™ . Female BALB/cJ mice aged 6 to 8 weeks were vaccinated with an emulsion containing 100 μg of recombinant mouse α-lactalbumin in an aqueous phase emulsified with 200 μg of Zymosan in 100 μL of IFA or 100 μL of MONTANIDE ™ . Four weeks after vaccination, the splenocyte frequencies of pro-inflammatory type 1 (IFNγ) and type 17 (IL-17) T cells were determined by ELISPOT analysis. Data represent the mean spot forming unit (SFU