[go: up one dir, main page]

WO2021084552A1 - Mimétique de mycobacterium tuberculosis pour l'immunisation et l'amélioration de l'efficacité d'un vaccin bcg - Google Patents

Mimétique de mycobacterium tuberculosis pour l'immunisation et l'amélioration de l'efficacité d'un vaccin bcg Download PDF

Info

Publication number
WO2021084552A1
WO2021084552A1 PCT/IN2020/050896 IN2020050896W WO2021084552A1 WO 2021084552 A1 WO2021084552 A1 WO 2021084552A1 IN 2020050896 W IN2020050896 W IN 2020050896W WO 2021084552 A1 WO2021084552 A1 WO 2021084552A1
Authority
WO
WIPO (PCT)
Prior art keywords
seq
vaccine
ligand
peptide fragment
sequence
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/IN2020/050896
Other languages
English (en)
Inventor
Ved Prakash DWIVEDI
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
International Centre for Genetic Engineering and Biotechnology
Original Assignee
International Centre for Genetic Engineering and Biotechnology
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by International Centre for Genetic Engineering and Biotechnology filed Critical International Centre for Genetic Engineering and Biotechnology
Publication of WO2021084552A1 publication Critical patent/WO2021084552A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/02—Bacterial antigens
    • A61K39/04—Mycobacterium, e.g. Mycobacterium tuberculosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • A61P31/06—Antibacterial agents for tuberculosis
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
    • C07K14/35—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Mycobacteriaceae (F)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/545—Medicinal preparations containing antigens or antibodies characterised by the dose, timing or administration schedule

Definitions

  • the present invention relates generally to the field of vaccines.
  • the present invention provides M.tb mimic useful in enhancing BCG vaccine efficacy or as stand-alone vaccine.
  • M.tb Mycobacterium tuberculosis
  • TB Tuberculosis
  • WHO 2017 the causative agent of Tuberculosis
  • M.tb infections may produce varied responses in individuals, ranging from asymptomatic infections to progressive pulmonary to extra-pulmonary TB and even death (Sharma et al., Challenges in the diagnosis & treatment of military tuberculosis, The Indian Journal of Medical Research, 2012, 135, 703-730).
  • the rate of progressive in the severity of TB depends upon the status of the host immune system.
  • BCG Mycobacterium bovis Bacillus Calmette-Guerin
  • BCG's limited vaccine efficacy is majorly attributed to its failure to induce a significant population of central memory T-cells (Bhattacharya et al., 2014a; Maggioli et al., Increased TNF-alpha/IFN- gamma/IL-2 and decreased TNF-alpha/IFN-gamma production by central memory T cells are associated with protective responses against Bovine Tuberculosis following BCG vaccination, Frontiers in Immunology, 2016, 7, 421; Singh et al., 2016; Vogelzang et al., Central memory CD+ T cells are responsible for the recombinant Bacillus Calmette-Guerin DelatureC::hly vaccine's superior protection against tuberculosis, the Journal of Infectious Diseases, 2014, 210, 1928- 1937) as animal modes vaccinated with BCG primarily develop antigen-specific CD4 + effector memory T-cells. Considering the lags in BCG immunization and increased global TB burden, it is crucial
  • composition comprising: (a) at least one peptide fragment having at least 80% sequence homology to at least one peptide fragment selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, and SEQ ID NO: 7; (b) at least one TLR1/2 ligand; and (c) at least one TLR9 ligand.
  • a vaccine comprising a liposomal composition
  • a liposomal composition comprising: (a) self-assembling lipids which form a liposome; (b) at least one peptide fragment having at least 80% sequence homology to at least one peptide fragment selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, and SEQ ID NO: 7; (c) at least one TLR1/2 ligand; and (d) at least one TLR9 ligand, wherein the said at least one peptide fragment, at least one TLR1/2 ligand, and at least one TLR9 ligand is liposomally bound.
  • a concomitant vaccine comprising: (a) BCG; and (b) a liposomal composition comprising: (i) self-assembling lipids which form a liposome; (ii) at least one peptide fragment having at least 80% sequence homology to at least one peptide fragment selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, and SEQ ID NO: 7; (iii) at least one TLR1/2 ligand; and (iv) at least one TLR9 ligand, wherein the said at least one peptide fragment, at least one TLR1/2 ligand, and at least one TLR9 ligand is liposomally bound.
  • a method of enhancing BCG vaccine efficacy comprising: concomitant subcutaneous administration of BCG vaccine and intranasal administration of a vaccine comprising a liposomal composition comprising: (a) self assembling lipids which form a liposome; (b) at least one peptide fragment having at least 80% sequence homology to at least one peptide fragment selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, and SEQ ID NO: 7; (c) at least one TLR1/2 ligand; and (d) at least one TLR9 ligand, wherein the said at least one peptide fragment, at least one TLR1/2 ligand, and at least one TLR9 ligand is liposomally bound.
  • a method of immunization comprising: (a) a first immunization comprising concomitant subcutaneous administration of BCG vaccine and intranasal administration of a vaccine comprising a liposomal composition comprising: (i) self assembling lipids which form a liposome; (ii) at least one peptide fragment having at least 80% sequence homology to at least one peptide fragment selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, and SEQ ID NO: 7; (iii) at least one TLR1/2 ligand; and (iv) at least one TLR9 ligand, wherein the said at least one peptide fragment, at least one TLR1/2 ligand, and at least one TLR9 ligand is liposomally bound; and (b) at least one subsequent booster immunization comprising concomitant subcutaneous administration of
  • Fig. 1 depicts the schematic diagram to show the preparation of M.tb mimic in accordance with an embodiment of the present invention.
  • Fig. 2 depicts the layout showing the experimental plan wherein naive Balb/c mice or mice vaccinated with BCG /M.tb mimic or with combination of BCG and M.tb mimic were challenged with H37Rv via the aerosol route with low-dose inoculum o about 220CFU/mice. Mice were sacrificed at various time points and lungs, spleen, and liver were harvested to look at the bacterial burden as well as profiling of immune responses, in accordance with an embodiment of the present invention.
  • Fig. 3 depicts (a) bar diagram to show the number of granulomas in all experimental groups; (b) CFU from lung; (c) CFU from spleen, (d) CFY from liver homogenates at 50 days post infection, in accordance with an embodiment of the present invention.
  • Fig. 4 depicts mice co-immunized with M.tb mimic and BCG infected with H37Rv M.tb followed by treatment with DOTS for 16 weeks. After 30 days of rest, these mice were treated with dexamethasone for 30 days followed by one more period of rest for 30 days. Mice were then sacrificed for CFU estimation to determine the rate of relapse post-treatment, in accordance with an embodiment of the present invention.
  • Fig. 5 depicts the determination of reactivation rate of latent M.tb.
  • Fig. 6a-d depicts the intracellular levels of innate cytokines in spleen; (a) IL-Ib; (b) IL-6; (c) TNF- a; (d) IL-10, in accordance with an embodiment of the present invention.
  • Fig. 7a-d depicts the intracellular levels of innate cytokines in lung; (a) IL-Ib; (b) IL-6; (c) TNF-a; (d) IL-10, in accordance with an embodiment of the present invention.
  • SEQ ID NO: 1-7 depicts the sequence of M.tb antigenic peptides SEQ ID NO: 1 (AWGRRLMIGTAAAVVLPG)
  • SEQ ID NO: 8 depicts the nucleotide sequence of a class A CpG ODN TLR9 ligand (ggGGTCAACGTTGAgggggg)
  • concomitant used herein refers to administration of a first and second vaccine component (BCG vaccine, M.tb mimic vaccine) at the same time (one after the other, on the same day, etc.).
  • the present invention provides a composition comprising: (a) at least one peptide fragment having at least 80% sequence homology to at least one peptide fragment selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO:
  • a composition as described herein wherein said TLR9 ligand is a class A ligand.
  • said TLR9 ligand is class B ligand.
  • said TLR9 ligand is a class C ligand.
  • said TLR9 ligand is class A ligand.
  • said TLR9 ligand is CpG ODN ligand having at least 80% sequence homology to a sequence as set forth in SEQ ID NO: 8.
  • said TLR9 ligand is CpG ODN ligand having sequence as set forth in SEQ ID NO: 8.
  • composition as described herein, wherein said TLR1/2 ligand is Pam 3 CSI ⁇ 4 .3HCI.
  • compositions as described herein wherein said composition comprises a first peptide fragment having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence homology to SEQ ID NO: 1, a second peptide fragment having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence homology to SEQ ID NO: 2, a third peptide fragment having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence homology to SEQ ID NO: 2,
  • said composition comprises a first peptide fragment having sequence as set forth in SEQ ID NO: 1, a second peptide fragment having sequence as set forth in SEQ ID NO: 2, a third peptide fragment having sequence as set forth in SEQ ID NO: 3, a fourth peptide fragment having sequence as set forth in SEQ ID NO: 4, a fifth peptide fragment having sequence as set forth in SEQ ID NO: 5, a sixth peptide fragment having sequence as set forth in SEQ ID NO: 6, and a seventh peptide fragment having sequence as set forth in SEQ ID NO: 7.
  • a composition comprising: a first peptide fragment having sequence as set forth in SEQ ID NO: 1, a second peptide fragment having sequence as set forth in SEQ ID NO: 2, a third peptide fragment having sequence as set forth in SEQ ID NO: 3, a fourth peptide fragment having sequence as set forth in SEQ ID NO: 4, a fifth peptide fragment having sequence as set forth in SEQ ID NO: 5, a sixth peptide fragment having sequence as set forth in SEQ ID NO: 6, a seventh peptide fragment having sequence as set forth in SEQ ID NO: 7, TLR9 ligand that is CpG ODN ligand having sequence as set forth in SEQ ID NO: 8, and TLR1/2 ligand that is Pam 3 CSK 4 .3HCI.
  • the present invention provides a vaccine comprising a liposomal composition, said liposomal composition comprising: (a) self-assembling lipids which form a liposome; and (b) a composition comprising: (i) at least one peptide fragment having at least 80% sequence homology to at least one peptide fragment selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, and SEQ ID NO: 7; (ii) at least one TLR1/2 ligand; and (iii) at least one TLR9 ligand, wherein the said at least one peptide fragment, at least one TLR1/2 ligand, and at least one TLR9 ligand is liposomally bound.
  • a vaccine composition as described herein wherein said liposome comprises cholesterol, L-a-phosphatidylcholine, and stearylamine.
  • said liposome is cationic.
  • the ratio of molal concentration of cholesterol to stearylamine is in the range of 1:1-1:5, preferably 1:2.
  • the ratio of molal concentration of cholesterol to L-a-phosphatidylcholine is in the range of 1:5-1:10, preferably 1:7.
  • the ratio of molal concentration of stearylamine to L-a- phosphatidylcholine is in the range of 1:1-1:5, preferably 1:3.5.
  • a vaccine composition as described herein, wherein said composition comprises a first peptide fragment having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence homology to SEQ ID NO: 1, a second peptide fragment having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence homology to SEQ ID NO: 2, a third peptide fragment having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence homology to SEQ ID
  • said composition comprises a first peptide fragment having sequence as set forth in SEQ ID NO: 1, a second peptide fragment having sequence as set forth in SEQ ID NO: 2, a third peptide fragment having sequence as set forth in SEQ ID NO: 3, a fourth peptide fragment having sequence as set forth in SEQ ID NO: 4, a fifth peptide fragment having sequence as set forth in SEQ ID NO: 5, a sixth peptide fragment having sequence as set forth in SEQ ID NO: 6, and a seventh peptide fragment having sequence as set forth in SEQ ID NO: 7.
  • a vaccine composition as described herein, wherein in said composition TLR9 ligand is a class A ligand.
  • said TLR9 ligand is class B ligand.
  • said TLR9 ligand is a class C ligand.
  • said TLR9 ligand is class A ligand.
  • said TLR9 ligand is CpG ODN ligand having at least 80% sequence homology to a sequence as set forth in SEQ ID NO: 8.
  • said TLR9 ligand is CpG ODN ligand having sequence as set forth in SEQ ID NO: 8.
  • a vaccine composition as described herein, wherein in said composition said TLR1/2 ligand is Pam 3 CSI ⁇ 4 .3HCI.
  • a vaccine composition as described herein wherein said vaccine is in lyophilized form. In another embodiment, the vaccine composition is in liquid form. In an embodiment, there is provided a vaccine composition as described herein, wherein said vaccine further comprises suitable adjuvants. In an embodiment, the vaccine is substantially adjuvant free.
  • a vaccine composition comprising: liposomes comprising cholesterol, L-a-phosphatidylcholine, and stearylamine having molal concentration ratio of 1:7:2, a first peptide fragment having sequence as set forth in SEQ ID NO: 1, a second peptide fragment having sequence as set forth in SEQ ID NO: 2, a third peptide fragment having sequence as set forth in SEQ ID NO: 3, a fourth peptide fragment having sequence as set forth in SEQ ID NO: 4, a fifth peptide fragment having sequence as set forth in SEQ ID NO: 5, a sixth peptide fragment having sequence as set forth in SEQ ID NO: 6, a seventh peptide fragment having sequence as set forth in SEQ ID NO: 7, TLR9 ligand that is CpG ODN ligand having sequence as set forth in SEQ ID NO: 8, and TLR1/2 ligand that is Pam 3 CSI ⁇ 4 .3HCI, and wherein the said a peptide fragments, TLR1/2 lig
  • the present invention provides a concomitant vaccine comprising: (a) BCG; and (b) a liposomal composition comprising: (i) self-assembling lipids which form a liposome; (ii) at least one peptide fragment having at least 80% sequence homology to at least one peptide fragment selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, and SEQ ID NO: 7; (iii) at least one TLR1/2 ligand; and (iv) at least one TLR9 ligand, wherein the said at least one peptide fragment, at least one TLR1/2 ligand, and at least one TLR9 ligand is liposomally bound.
  • BCG is in attenuated form.
  • BCG is obtained from attenuated live bovine tuberculosis bacillus, Mycobacterium bovis.
  • any conventional strain of bovine tuberculosis bacillus may be used.
  • a concomitant vaccine as described herein wherein said liposome comprises cholesterol, L-a-phosphatidylcholine, and stearylamine.
  • said liposome is cationic.
  • the ratio of molal concentration of cholesterol to stearylamine is in the range of 1:1-1:5, preferably 1:2.
  • the ratio of molal concentration of cholesterol to L-a-phosphatidylcholine is in the range of 1:5-1:10, preferably 1:7.
  • the ratio of molal concentration of stearylamine to L-a- phosphatidylcholine is in the range of 1:1-1:5, preferably 1:3.5.
  • a concomitant vaccine as described herein, wherein said vaccine comprises a first peptide fragment having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence homology to SEQ ID NO: 1, a second peptide fragment having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence homology to SEQ ID NO: 2, a third peptide fragment having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence homology to
  • said vaccine comprises a first peptide fragment having sequence as set forth in SEQ ID NO: 1, a second peptide fragment having sequence as set forth in SEQ ID NO: 2, a third peptide fragment having sequence as set forth in SEQ ID NO: 3, a fourth peptide fragment having sequence as set forth in SEQ ID NO: 4, a fifth peptide fragment having sequence as set forth in SEQ ID NO: 5, a sixth peptide fragment having sequence as set forth in SEQ ID NO: 6, and a seventh peptide fragment having sequence as set forth in SEQ ID NO: 7.
  • TLR9 ligand is a class A ligand.
  • said TLR9 ligand is class B ligand.
  • said TLR9 ligand is a class C ligand.
  • said TLR9 ligand is class A ligand.
  • said TLR9 ligand is CpG ODN ligand having at least 80% sequence homology to a sequence as set forth in SEQ ID NO: 8.
  • said TLR9 ligand is CpG ODN ligand having sequence as set forth in SEQ ID NO: 8.
  • TLR1/2 ligand is Pam 3 CSK .3HCI.
  • a concomitant vaccine as described herein, wherein said vaccine is in lyophilized form.
  • the vaccine is in liquid form.
  • the vaccine is a combination of lyophilized form and liquid form.
  • a concomitant vaccine as described herein, wherein said vaccine further comprises suitable adjuvants.
  • the vaccine is substantially adjuvant free.
  • a concomitant vaccine comprising: live attenuated BCG obtained from bovine tuberculosis bacillus, Mycobacterium bovis, liposomes comprising cholesterol, L-a-phosphatidylcholine, and stearylamine having molal concentration ratio of 1:7:2, a first peptide fragment having sequence as set forth in SEQ ID NO: 1, a second peptide fragment having sequence as set forth in SEQ ID NO: 2, a third peptide fragment having sequence as set forth in SEQ ID NO: 3, a fourth peptide fragment having sequence as set forth in SEQ ID NO: 4, a fifth peptide fragment having sequence as set forth in SEQ ID NO: 5, a sixth peptide fragment having sequence as set forth in SEQ ID NO: 6, a seventh peptide fragment having sequence as set forth in SEQ ID NO: 7, TLR9 ligand that is CpG ODN ligand having sequence as set forth in SEQ ID NO: 8, and TLR1/2 ligand
  • the present invention provides a method of enhancing BCG vaccine efficacy, said method comprising: concomitant subcutaneous administration of BCG vaccine and intranasal administration of a vaccine comprising a liposomal composition comprising: (a) self-assembling lipids which form a liposome; (b) at least one peptide fragment having at least 80% sequence homology to at least one peptide fragment selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, and SEQ ID NO: 7; (c) at least one TLR1/2 ligand; and (d) at least one TLR9 ligand, wherein the said at least one peptide fragment, at least one TLR1/2 ligand, and at least one TLR9 ligand is liposomally bound.
  • BCG is obtained from attenuated live bovine tuberculosis bacillus, Mycobacterium bovis.
  • any conventional strain of bovine tuberculosis bacillus may be used.
  • a method of enhancing BCG vaccine efficacy as described herein wherein said liposome comprises cholesterol, L-a-phosphatidylcholine, and stearylamine.
  • said liposome is cationic.
  • the ratio of molal concentration of cholesterol to stearylamine is in the range of 1:1-1:5, preferably 1:2.
  • the ratio of molal concentration of cholesterol to L-a-phosphatidylcholine is in the range of 1:5-1:10, preferably 1:7.
  • the ratio of molal concentration of stearylamine to L-a- phosphatidylcholine is in the range of 1:1-1:5, preferably 1:3.5.
  • a method of enhancing BCG vaccine efficacy as described herein wherein said liposomal composition comprises a first peptide fragment having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence homology to SEQ ID NO: 1, a second peptide fragment having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence homology to SEQ ID NO: 2, a third peptide fragment having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 96%,
  • said liposomal composition comprises a first peptide fragment having sequence as set forth in SEQ ID NO: 1, a second peptide fragment having sequence as set forth in SEQ ID NO: 2, a third peptide fragment having sequence as set forth in SEQ ID NO: 3, a fourth peptide fragment having sequence as set forth in SEQ ID NO: 4, a fifth peptide fragment having sequence as set forth in SEQ ID NO: 5, a sixth peptide fragment having sequence as set forth in SEQ ID NO: 6, and a seventh peptide fragment having sequence as set forth in SEQ ID NO: 7.
  • TLR9 ligand is a class A ligand.
  • said TLR9 ligand is class B ligand.
  • said TLR9 ligand is a class C ligand.
  • said TLR9 ligand is class A ligand.
  • said TLR9 ligand is CpG ODN ligand having at least 80% sequence homology to a sequence as set forth in SEQ ID NO: 8.
  • said TLR9 ligand is CpG ODN ligand having sequence as set forth in SEQ ID NO: 8.
  • TLR1/2 ligand is Pam 3 CSI ⁇ 4 .3HCI.
  • the vaccine is substantially adjuvant free.
  • BCG vaccine comprising live attenuated BCG obtained from bovine tuberculosis bacillus, Mycobacterium bovis, is administered subcutaneously and concomitantly intranasal administration of a vaccine comprising liposomes comprising cholesterol, L-a- phosphatidylcholine, and stearylamine having molal concentration ratio of 1:7:2, a first peptide fragment having sequence as set forth in SEQ ID NO: 1, a second peptide fragment having sequence as set forth in SEQ ID NO: 2, a third peptide fragment having sequence as set forth in SEQ ID NO: 3, a fourth peptide fragment having sequence as set forth in SEQ ID NO: 4, a fifth peptide fragment having sequence as set forth in SEQ ID NO: 5, a sixth peptide fragment having sequence as set forth in SEQ ID NO: 6, a seventh peptide fragment having sequence as set forth in SEQ ID
  • the present invention provides a method of immunization comprising: (a) a first immunization comprising concomitant subcutaneous administration of BCG vaccine and intranasal administration of a vaccine comprising a liposomal composition comprising: (i) self-assembling lipids which form a liposome; (ii) at least one peptide fragment having at least 80% sequence homology to at least one peptide fragment selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, and SEQ ID NO: 7; (iii) at least one TLR1/2 ligand; and (iv) at least one TLR9 ligand, wherein the said at least one peptide fragment, at least one TLR1/2 ligand, and at least one TLR9 ligand is liposomally bound; and (b) at least one subsequent booster immunization comprising concomitant subcutaneous administration of BCG vaccine and intran
  • total peptide dosage for human administration in first administration is in the range of 0.01- lmg/kg, preferably in the range of 0.04-0.08mg/kg, more preferably about 0.06mg/kg.
  • total peptide dosage for human administration in booster administration is in the range of 0.01- lmg/kg, preferably in the range of 0.04-0.08mg/kg, more preferably about 0.06mg/kg.
  • total TLR1/2 ligand dosage for human administration in first administration is in the range of 0.001-0. lmg/kg, preferably in the range of 0.005-0. lmg/kg, more preferably about 0.009mg/kg.
  • total TLR9 ligand dosage for human administration in first administration is in the range of 0.0001-0. Olmg/kg, preferably in the range of 0.0009-0. Olmg/kg, more preferably about O.OOlmg/kg.
  • a vaccine comprising a liposomal composition as described herein, for use in immunization against TB.
  • Mycobacterium tuberculosis H37Rv and BCG cultures were grown in 7H9 (Middlebrooks, Difco, USA) medium supplemented with 10% OADC (oleic acid, albumin, dextrose and catalase; Difco, USA) and with0.05% Tween 80 and 0.5% glycerol, and cultures were grown to mid-log phase. Aliquots of the cultures in 20% glycerol were preserved at -80°C and cryopreserved stocks were used for infections.
  • 7H9 Middlebrooks, Difco, USA
  • OADC oleic acid, albumin, dextrose and catalase
  • Tween 80 and 0.5% glycerol oleic acid, albumin, dextrose and catalase
  • mice were infected with H37Rv via the aerosol route using a Madison aerosol chamber (University of Wisconsin, Madison, Wl, USA) with its nebulizer pre-calibrated to deposit around of 220 bacilli to the lungs of each mouse (Chaterjee et al., 2011). Briefly, bacterial stocks were recovered from freezer and quickly thawed and subjected to light ultra-sonication to obtain a single cell suspension. 15ml of the bacterial cell suspension (lOxlO 16 cells per ml) was placed in the nebulizer to deliver the desired number of CFUs to the lungs of animals placed inside the chamber.
  • mice At day one post infection, three randomly selected mice sacrificed and organs were harvested, homogenized in 0.2pm filtered PBS and plated onto 7H11 Middlebrooks (Difco, USA) plates containing 10% oleic acid, albumin, dextrose and catalase. Undiluted, 10-fold diluted, and 100-fold diluted lung, liver, and spleen cells homogenates were plated in triplicate on the 7H11 plates and incubated at 37°C for 21-28 days. Colonies were counted and CFU was calculated accordingly.
  • mice infected with M.tb with low-dose aerosol infection model were treated with lOmg/kg of 1NH and RIF administered ad libitum (in the drinking water) or treated to mice co-immunized with BCG and M.tb mimic or immunized with BCG alone for 12 weeks starting at 4 th week after infection. These mice were then rested for 30 days followed by treatment with dexamethasone (5mg/kg administered intraperitoneally) three times per week for 30 days. 10 mice from each group were then sacrificed and CFUs were estimated from lung homogenates to determine the reactivation rate of M.tb.
  • Peptide screening was done on the basis of their capacity to induce M.tb specific T- cell activation and high level of IFN-gamma production.
  • mice were infected with H37Rv strain of M.tb and were subjected to 45 days of DOTS therapy 15 days post infection. The mice were rested for 30 days post-therapy. T-cells from the infected mice as well as DOTS treated mice were isolated and co-cultured with dendritic cells, which were pulsed with T-cell epitopes/peptides. Following which peptide-specific T cell (subsets of CD4 + and CD8 + T cells) activation and proliferation was observed. To investigate the role of these peptides in T-cell activation, experiments were performed using individual as well as pooled peptides, and complete soluble antigen (CSA) of M.tb as a control.
  • CSA complete soluble antigen
  • the first immunization dosage comprises 800pg of each peptide per kg of animal weight, 120pg of TLR1/2 ligand per kg of animal weight, and 20pg of TLR9 ligand per kg of animal weight contained in the liposomes.
  • the dosage comprises 40opg of each peptide per kg of animal weight, 120pg ofTLRl/2 ligand per kg of animal weight, and 20pg ofTLR9 ligand per kg of animal weight contained in the liposomes.
  • mice All animals [Balb/c and C57BL.6 mice (6-8 weeks of age) ⁇ were maintained in ICGEB animal facility.
  • mice were immunized with (i) BCG (subcutaneous) (10 6 bacteria), (ii) M.tb mimics (intranasal); (iii) BCG + M.tb mimics, and (iv) vector alone.
  • BCG vaccinated and mimic immunized mice were boosted with once-a-week, three week-long M.tb mimic boosting regimen.
  • Mice were subsequently rested for 20 days and then challenged with M.tb strain H37Rv by aerosol route ( ⁇ 220CFU/mouse). Organs like lungs, livers, and spleens were harvested for determination of bacterial burden.
  • Fig. 2 depicts the experimental layout.
  • Fig. 3a depicts the granuloma score.
  • Fig. 3b-d depicts the CFU count from lung tissue, spleen tissue, and liver tissue sample respectively. It can be seen that for the combination of BCG and M.tb mimics, there is a drastic drop in CFU count across all 3 different samples compared to treatment with BCG or M.tb mimics alone.
  • M.tb mimic immunization protects animals against TB recurrence due to relapse
  • mice were co-immunized with M.tb mimic and BCG (BCG vaccinated and mimic immunized mice were boosted with once-a-week, three week-long M.tb mimic boosting regimen) and infected with H37Rv M.tb followed by treatment with DOTS for 16 weeks. After 30 days of rest, the mice were treated with dexamethasone for 30 days followed by one more period of rest for 30 days. Mice were then sacrificed for CFU estimation to determine the rate of relapse post-treatment.
  • Fig. 4 depicts the treatment paradigm.
  • Fig. 5 depicts the reactivation rate of latent M.tb in three different treatment paradigms. As seen in Fig. 5, while in control (no vaccine), relapse rate was about 70%, in BCG vaccine alone, relapse rate was about 45%, while in mice treated with BCG + M.tb mimics, the relapse rate was about 20%. These data clearly show that the concomitant administration of M.tb mimics of the present invention along with BCG significantly decreases TB reactivation rate compared to BCG administration alone.
  • M.tb mimic immunization induces protective cytokine response
  • M.tb mimic co-immunization (BCG vaccinated and mimic immunized mice were boosted with once-a-week, three week-long M.tb mimic boosting regimen) enhances IL-Ib, IL-6, and TNF-a producing cells (Fig. 6a-c), with no significant decrease in percentage of IL-10 producing cells (Fig. 5d) in spleen. A similar profile was observed in the lungs of co-immunized mice (Fig. 7a-d). These data suggest that M.tb mimic co-immunization enhances antigen specific Thl and Thl7 responses, which impart protection against TB.
  • M.tb co-immunized mice also show a significant increase in the number of CD4 + CD8 + double positive T-cells as compared to mice immunized with BCG alone (data not shown). These double positive T-cells have been reported to mount a strong protective response in various disease settings (Overgaard et al., 2015). Overall, these data collective show that the M.tb mimic of the present invention when administered (alone) to animals, provides protection against TB with efficiency similar to that of BCG vaccine alone.
  • the M.tb mimic enhances the efficacy of BCG by not only reducing bacterial burden but also enabling the host system in mounting a potent immune response by activating protective T-cells along with an enriched central memory T-cell repertoire, thus increasing the efficacy of existing TB vaccine.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Organic Chemistry (AREA)
  • Communicable Diseases (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Pulmonology (AREA)
  • Genetics & Genomics (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Immunology (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Molecular Biology (AREA)
  • Epidemiology (AREA)
  • Biophysics (AREA)
  • Biochemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Oncology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Peptides Or Proteins (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Medicinal Preparation (AREA)

Abstract

La présente invention concerne de nouveaux fragments peptidiques liés par liposome complexés avec un ou plusieurs ligands TLR1/2 et TLR9 utiles en tant que vaccin contre la tuberculose et pour améliorer l'efficacité du vaccin BCG lors d'une utilisation concomitante.
PCT/IN2020/050896 2019-10-30 2020-10-21 Mimétique de mycobacterium tuberculosis pour l'immunisation et l'amélioration de l'efficacité d'un vaccin bcg Ceased WO2021084552A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
IN201911043992 2019-10-30
IN201911043992 2019-10-30

Publications (1)

Publication Number Publication Date
WO2021084552A1 true WO2021084552A1 (fr) 2021-05-06

Family

ID=75715847

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/IN2020/050896 Ceased WO2021084552A1 (fr) 2019-10-30 2020-10-21 Mimétique de mycobacterium tuberculosis pour l'immunisation et l'amélioration de l'efficacité d'un vaccin bcg

Country Status (1)

Country Link
WO (1) WO2021084552A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116807974A (zh) * 2023-06-05 2023-09-29 四川大学 含载药脂质体和卡介苗的复合物递药系统制备方法和应用

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2007063421A2 (fr) * 2005-12-01 2007-06-07 The Provost, Fellows And Scholars Of The College Of The Holy And Undivided Trinity Of Queen Elizabeth, Near Dublin Compositions et methodes relatives au traitement du cancer et de maladies infectieuses
WO2009123480A1 (fr) * 2008-04-01 2009-10-08 Virionyx Corporation Ltd Agents anti-infectieux et utilisations de ces derniers
US20110236411A1 (en) * 2007-09-27 2011-09-29 Dako Denmark A/S MHC Multimers in Tuberculosis Diagnostics, Vaccine and Therapeutics

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2007063421A2 (fr) * 2005-12-01 2007-06-07 The Provost, Fellows And Scholars Of The College Of The Holy And Undivided Trinity Of Queen Elizabeth, Near Dublin Compositions et methodes relatives au traitement du cancer et de maladies infectieuses
US20110236411A1 (en) * 2007-09-27 2011-09-29 Dako Denmark A/S MHC Multimers in Tuberculosis Diagnostics, Vaccine and Therapeutics
WO2009123480A1 (fr) * 2008-04-01 2009-10-08 Virionyx Corporation Ltd Agents anti-infectieux et utilisations de ces derniers

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
ANDERSEN ET AL.: "Recall of long-lived immunity to Mycobacterium tuberculosis infection in mice", JOURNAL OF IMMUNOLOGY, vol. 154, no. 7, 1 April 1995 (1995-04-01), Baltimore, Md. : 1950, pages 3359 - 72, XP001131906 *
MAEYAMA ET AL.: "The mucosal adjuvanticity of the oligodeoxynucleotides containing a non-methylated CpG motif on BCG and diphtheria toxoid", VACCINE, vol. 27, no. 8, 9 January 2009 (2009-01-09), pages 1166 - 73, XP025924624, DOI: 10.1016/j.vaccine.2008.12.025 *
MUSTAFA ET AL.: "Identification and HLA restriction of naturally derived Thl- cell epitopes from the secreted Mycobacterium tuberculosis antigen 85B recognized by antigen-specific human CD 4(+) T- cell lines", INFECTION AND IMMUNITY, vol. 68, no. 7, 1 July 2000 (2000-07-01), pages 3933 - 40, XP055022880 *
VAN DISSEL ET AL.: "Ag85B-ESAT-6 adjuvanted with IC31@ promotes strong and long-lived Mycobacterium tuberculosis specific T cell responses in volunteers with previous BCG vaccination or tuberculosis infection", VACCINE, vol. 29, no. 11, 24 December 2010 (2010-12-24), pages 2100 - 9, XP028152756 *
WEDLOCK ET AL.: "Enhanced protection against bovine tuberculosis after coadministration of Mycobacterium bovis BCG with a Mycobacterial protein vaccine-adjuvant combination but not after coadministration of adjuvant alone", CLINICAL AND VACCINE IMMUNOLOGY, vol. 15, no. 5, 5 May 2008 (2008-05-05), pages 765 - 72, XP055821847 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116807974A (zh) * 2023-06-05 2023-09-29 四川大学 含载药脂质体和卡介苗的复合物递药系统制备方法和应用

Similar Documents

Publication Publication Date Title
Nieuwenhuizen et al. Next-generation vaccines based on Bacille Calmette–Guérin
Gong et al. The current status, challenges, and future developments of new tuberculosis vaccines
US8741313B2 (en) Polypeptide vaccine and vaccination strategy against mycobacterium
US11224646B2 (en) Intranasal delivery of a cyclic-di-nucleotide adjuvanted vaccine for tuberculosis
Yuan et al. Immunogenicity and protective efficacy of a tuberculosis DNA vaccine expressing a fusion protein of Ag85B-Esat6-HspX in mice
USRE46583E1 (en) Vaccines comprising TB10.4
Commandeur et al. The in vivo expressed Mycobacterium tuberculosis (IVE-TB) antigen Rv2034 induces CD4+ T-cells that protect against pulmonary infection in HLA-DR transgenic mice and guinea pigs
Romano et al. Immunogenicity and protective efficacy of tuberculosis subunit vaccines expressing PPE44 (Rv2770c)
Singh et al. Manipulation of BCG vaccine: a double-edged sword
Sable et al. Multicomponent antituberculous subunit vaccine based on immunodominant antigens of Mycobacterium tuberculosis
Fan et al. Differential immunogenicity and protective efficacy of DNA vaccines expressing proteins of Mycobacterium tuberculosis in a mouse model
Kohama et al. Mucosal immunization with recombinant heparin-binding haemagglutinin adhesin suppresses extrapulmonary dissemination of Mycobacterium bovis bacillus Calmette-Guerin (BCG) in infected mice
JP2010508254A (ja) 結核の予防ワクチン
Yin et al. A promising listeria-vectored vaccine induces Th1-type immune responses and confers protection against tuberculosis
WO2021084552A1 (fr) Mimétique de mycobacterium tuberculosis pour l'immunisation et l'amélioration de l'efficacité d'un vaccin bcg
Elhay et al. Immunological requirements for a subunit vaccine against tuberculosis
Peeridogaheh et al. Evaluation of immune responses to a DNA vaccine encoding Ag85a-Cfp10 antigen of Mycobacterium tuberculosis in an animal model
Siddiqui et al. Prime-boost vaccination strategy with bacillus Calmette–Guérin (BCG) and liposomized alpha-crystalline protein 1 reinvigorates BCG potency
Hernandez-Pando et al. Recombinant BCG vaccine candidates
Safar et al. The effect of delivery systems on the induction of T helper 1 cell response to an ESAT6-like protein Rv3619c and identification of its immunodominant peptides
Ji et al. Boosting BCG-primed mice with chimeric DNA vaccine HG856A induces potent multifunctional T cell responses and enhanced protection against Mycobacterium tuberculosis
JP2001518781A (ja) マイコバクテリウム組換えワクチン
Gillard et al. Functional programming of innate immune cells in response to bordetella pertussis infection and vaccination
Horwitz et al. Novel vaccines against tuberculosis
US20150224188A1 (en) Use of membrane vesicle-based vaccine against m. tuberculosis

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 20881589

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 20881589

Country of ref document: EP

Kind code of ref document: A1