Novel immunologic adjuvant, compound and application thereof
An immune adjuvant and compound technology, applied in the field of compounds and new immune adjuvants, can solve the problems of lack of immune cell activation enhancement, vegetable oil adjuvant and Freund's adjuvant local stimulation, etc., to enhance the induction ability and enhance the binding ability , Enhance the effect of TLR7 activation
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Embodiment 1
[0052]Embodiment 1 AlumT-I preparation:
[0053] Add 10 mg (14 μmol) of T-I to 2.5 mL of purified deionized water, and add 14 μL solution containing 0.56 mg NaOH to form a clear solution. Then add 2 mL of Alhydrogel (Invivogen, containing 9.0-11.0 mg / mL of aluminum), 1 mL of histidine buffer solution (100 mM, pH 6.5), and finally add 4.5 mL of deionized water. The resulting mixture was stirred slowly at room temperature for 1 hour to obtain AlumT-I adjuvant (about 10 mL). Store at room temperature 20-25°C. The mass ratio of T-I to aluminum is 1:2, and the molar ratio is 1:53.
[0054] The preparation of AlumT-II, AlumT-III, and AlumT-IV is the same as that of AlumT-I.
Embodiment 2
[0055] Example 2 Preparation of 54-peptide vaccine (Vac) with peptide 54-peptide in FZD7 protein as antigen
[0056] Frizzled7 (FZD7) is an antigen marker of tumor stem cells ["Frizzled7 as an emerging target for cancer therapy". -7as a potential therapeutic target incolorectal cancer".Neoplasia.2008Jul; 10(7):697-705], the main peptide 54 was selected as an antigen to prepare a therapeutic vaccine;
[0057] Add 10 mg of APVCTVLDQAIPPCRSLCERARQGCEALMNKFGFQWPERLRCENFPVHGAGEIC (54 peptide) to 10 mL of AlumT-I adjuvant, and shake gently at room temperature to obtain the 54 peptide vaccine (Vac1).
Embodiment 354
[0058] Example 3 Using 54 peptide vaccine (Vac1 vaccine, i.e. 54 peptide+AlumT-I) as the immune anti-tumor experiment of therapeutic vaccine
[0059] BALB / c mice aged 4-6 weeks, which are homologous to CT26.WT cells, were selected and subcutaneously injected with 2.0×10 5 CT26.WT cells, when the tumor diameter reached 4-5 mm, the tumor-bearing mice were randomly divided into PBS group, Alum group (Alhydrogel), Alum+54 peptide, T-I+54 peptide, Vac1 group (vaccine group, AlumT-I+54 peptide) group, 10 rats in each group. Start group injection and vaccine injection, Alum group contains Al (1mg / Alhydrogel 100μL), Alum+54 peptide group (containing Al 1mg / Alhydrogel 100μL plus 54 peptide 100μg), T-I+54 peptide group (containing T-I 100μg plus 54 peptide 100 μg plus PBS 100 μL), Vac1 group (vaccine group, AlumT-I 100 μL plus 54 peptide 100 μg). The injection volume of each group was 100 μL, and the vaccine was injected on the 8th day, the 12th day, the 15th day, the 19th day and the...
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