Glycopeptide vaccine
A peptide antigen, compound technology, applied in the field of vaccines to prevent or reduce the incidence of malaria in subjects, can solve problems such as low efficacy
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Embodiment 1
[0337] Example 1 - Synthesis of Peptides for Conjugation
[0338] 5-azidovaleryl-FFRK-AAAHSLSNVYDFNLLLERD (SEQ ID NO: 1-SEQ ID NO: 3)
[0339] 5-azidovaleryl-FFRK-AAAHSLSNVYDFNLLLERD (SEQ ID NO: 1 - SEQ ID NO: 3) by FmocSPPS in 4-(4-hydroxymethyl-3-methoxyphenoxy)butyric acid (HMPB) Synthesis on resin preloaded with Fmoc-L-aspartate 4-tert-butyl ester (Fmoc-Asp(tBu)) as the first amino acid. Alpha-amino acids with the following side chain protecting groups were used: Arg(Pbf), Lys(Boc), Ser(tBu), Asn(Trt), His(Trt), Tyr(tBu), Asp(tBu), Glu(tBu) ), and use Initiator+Alstra TMThe microwave peptide synthesizer synthesizes peptides on a 0.1 mmol scale. The resin was swollen in DMF (20 min, 70° C., 50 W), followed by a synthetic reaction cycle consisting of: Fmoc deprotection with 20% piperidine in DMF (3 min, then 10 min, room temperature); and amino acid coupling (5 min, 75°C, 50W) using 5 equiv of the protected amino acid (0.5M) in DMF via 5 equiv of diisopropylcarbodiimi...
Embodiment 2
[0345] Example 2: Synthesis of glycolipid-linker for oxime conjugation
[0346] 4-(N-((9H-Fluoren-9-yl)methoxycarbonyl)-L-valyl-L-citrullineamido)benzyl 4-nitrophenylcarbonate (Fmoc-VCPAB- pNP)
[0347]
[0348] To a mixture of the alcohol Fmoc-VC-PABOH (Dubowchik, 2002) (400 mg, 0.665 mmol) in DMF (6.0 mL) was added bis(4-nitrophenyl)carbonate (255 mg, 0.796 mmol) and i-Pr 2 NEt (0.13 mL, 0.75 mmol). After stirring under Ar at room temperature for 18 h, the solvent was co-evaporated several times with toluene in a rotary evaporator. Flash chromatography on silica gel (gradient elution, MeOH / CHCl 3 = 0:100 to 8:92) to give the title compound as a pale yellow solid (380 mg, 75%). 1 H NMR (500MHz, d 6 -DMSO)δ0.85(d,J=6.7Hz,3H),0.88(d,J=6.7Hz,3H),1.33-1.49(m,2H),1.56-1.64(m,1H),1.67-1.74 (m,1H),1.95-2.02(m,1H),2.91-2.97(m,1H),2.99-3.06(m,1H),3.92(dd,J=7.2,8.7Hz,1H),4.20-4.26 (m,2H),4.28-4.33(m,1H),4.39-4.43(m,1H),5.24(s,2H),5.39(br s,2H),5.98(br t,J=5.7Hz,1H ), 7.30-7....
Embodiment 3
[0358] Example 3: Synthesis of glycolipid-peptide conjugates
[0359] SPAAC conjugate vaccine
[0360] MaGC-PAB-CV-cyclooctyne and conjugate compound V.S.FFRK.OVA were synthesized as previously described LP (C 26 ). (Anderson RJ, 2017)
[0361]
[0362] Conjugate V.S.FFRK.NVY SP
[0363]
[0364] 5-azidovaleryl-FFRK-NVYDFNLL (SEQ ID NO: 1-SEQ ID NO: 2) (1.6 mg, 0.96 μmol) and MaGC-PABA-CV-cyclooctyne (1.0 mg, 0.69 μmol) were added The solution in DMSO (100 μL) was kept at room temperature for 2 days. The product solution was passed through semi-preparative HPLC (Phenomenex Luna C18(1), 4.6 μm, 250×10 mm, 40°C, mobile phase A=100:0.05 water / TFA; mobile phase B=100:0.05MeOH / TFA; gradient program : T0=80%B, T12=100%B, T14=100%B, T14.5=80%B, T16=80%B; flow rate: T0=3mL / min, T1=3mL / min, T12=4.5 mL / min, T14=4.5mL / min, T16=3mL / min), get V.S.FFRK.NVY SP , as a white powder (1.8 mg, 82%). HRMS-ESI m / z calculation C 162 H 257 N 27 O 35 [M+2H] 2+ 1570.4580, measured...
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