ABSTRACT
This study describes the design and synthesis of five TF-based cancer vaccine candidates using a lipid A mimetic as the carrier and a built-in adjuvant. All synthesized conjugates elicited robust and consistent TF-specific immune responses in mice without external adjuvants. Immunological studies subsequently conducted in wild-type and TLR4 knockout C57BL/6 mice demonstrated that the activation of TLR4 was the main reason that the synthesized lipid A mimetics increased the TF-specific immune responses. All antisera induced by these conjugates can specifically recognize, bind to, and induce the lysis of TF-positive cancer cells. Moreover, representative conjugates 2 and 3 could effectively reduce the growth of tumors and prolong the survival time of mice in vivo, and the efficacies were better than glycoprotein TF-CRM197 with alum adjuvant. Lipid A mimetics could therefore be a promising platform for the development of new carbohydrate-based vaccine carriers with self-adjuvanting properties for the treatment of cancer.
Subject(s)
Adjuvants, Immunologic , Cancer Vaccines , Drug Design , Lipid A , Mice, Inbred C57BL , Animals , Lipid A/analogs & derivatives , Lipid A/chemistry , Lipid A/pharmacology , Cancer Vaccines/immunology , Cancer Vaccines/pharmacology , Cancer Vaccines/chemical synthesis , Adjuvants, Immunologic/pharmacology , Adjuvants, Immunologic/chemical synthesis , Adjuvants, Immunologic/chemistry , Mice , Mice, Knockout , Humans , Female , Toll-Like Receptor 4/metabolism , Cell Line, TumorABSTRACT
Muramyl dipeptide (MDP) is the smallest essential peptidoglycan substructure capable of promoting both innate and adaptive immune responses. Herein, we report on the design, synthesis, and in vivo study of the adjuvant properties of two novel MDP analogs containing an achiral adamantyl moiety attached to the desmuramyl dipeptide (DMP) pharmacophore and additionally modified by one mannosyl subunit (derivative 7) or two mannosyl subunits (derivative 11). Mannose substructures were introduced in order to assess how the degree of mannosylation affects the immune response and nucleotide-binding oligomerization-domain-containing protein 2 (NOD2) binding affinity, compared to the reference compound ManAdDMP. Both mannosylated MDP analogs showed improved immunomodulating properties, while the di-mannosylated derivative 11 displayed the highest, statistically significant increase in anti-OVA IgG production. In this study, for the first time, the di-mannosylated DMP derivative was synthesized and immunologically evaluated. Derivative 11 stimulates a Th-2-polarized type of immune reaction, similar to the reference compound ManAdDMP and MDP. Molecular dynamics (MD) simulations demonstrate that 11 has a higher NOD2 binding affinity than 7, indicating that introducing the second mannose significantly contributes to the binding affinity. Mannose interacts with key amino acid residues from the LRR hydrophobic pocket of the NOD2 receptor and loop 2.
Subject(s)
Acetylmuramyl-Alanyl-Isoglutamine , Adamantane , Adjuvants, Immunologic , Mannose , Adamantane/chemistry , Adamantane/analogs & derivatives , Mannose/chemistry , Adjuvants, Immunologic/chemistry , Adjuvants, Immunologic/pharmacology , Adjuvants, Immunologic/chemical synthesis , Acetylmuramyl-Alanyl-Isoglutamine/chemistry , Acetylmuramyl-Alanyl-Isoglutamine/pharmacology , Animals , Molecular Dynamics Simulation , Mice , Nod2 Signaling Adaptor Protein/metabolism , Nod2 Signaling Adaptor Protein/chemistry , HumansABSTRACT
Toll-like receptor (TLR)-7 agonists are immunostimulatory vaccine adjuvants. A systematic structure-activity relationship (SAR) study of TLR7-active 1-benzyl-2-butyl-1H-imidazo[4,5-c]quinolin-4-amine led to the identification of a potent hTLR7-specific p-hydroxymethyl IMDQ 23 with an EC50 value of 0.22 µM. The SAR investigation also resulted in the identification of TLR7 selective carboxamide 12 with EC50 values of 0.32 µM for hTLR7 and 18.25 µM for hTLR8. In the vaccination study, TLR7-specific compound 23 alone or combined with alum (aluminum hydroxide wet gel) showed adjuvant activity for a spike protein immunogen in mice, with enhanced anti-spike antibody production. Interestingly, the adjuvant system comprising carboxamide 12 and alum showed prominent adjuvant activity with high levels of IgG1, IgG2b, and IgG2c in immunized mice, confirming a balanced Th1/Th2 response. In the absence of any apparent toxicity, the TLR7 selective agonists in combination with alum may make a suitable vaccine adjuvant.
Subject(s)
Adjuvants, Immunologic , Toll-Like Receptor 7 , Toll-Like Receptor 7/agonists , Structure-Activity Relationship , Animals , Humans , Adjuvants, Immunologic/pharmacology , Adjuvants, Immunologic/chemistry , Adjuvants, Immunologic/chemical synthesis , Mice , Female , Alum Compounds/pharmacology , Alum Compounds/chemistry , Mice, Inbred BALB C , Imidazoles/chemistry , Imidazoles/pharmacology , Imidazoles/chemical synthesisABSTRACT
TLR7/8 agonists are versatile immune stimulators capable of treating various diseases such as viral infections, autoimmune, and cancer. Despite the structural similarity of TLR7/8, their immune stimulation mechanisms and time-course responses significantly differ. In this study, a new series of TLR7-selective agonists was synthesized utilizing the economical building block 2,6-dichloropurine. Compound 27b showed the most potent activity on hTLR7 with an EC50 of 17.53 nM and demonstrated high hTLR7 selectivity (224 folds against TLR8). 27b effectively stimulated the secretion of proinflammatory cytokines in mouse macrophages and enhanced intranasal vaccine efficacy against influenza A virus in vivo. Assessment of humoral and mucosal antibody titers confirmed that 27b elevates IgG and IgA levels, protecting against both homologous and heterologous influenza viral infections. These findings suggest that 27b is a promising candidate as a vaccine adjuvant to prevent viral infections or as a robust immunomodulator with prolonged activity for treating immune-suppressed diseases.
Subject(s)
Administration, Intranasal , Drug Design , Influenza Vaccines , Purines , Toll-Like Receptor 7 , Toll-Like Receptor 7/agonists , Animals , Mice , Humans , Influenza Vaccines/immunology , Influenza Vaccines/administration & dosage , Purines/pharmacology , Purines/chemistry , Adjuvants, Vaccine/pharmacology , Adjuvants, Vaccine/chemistry , Structure-Activity Relationship , Mice, Inbred BALB C , Female , Orthomyxoviridae Infections/prevention & control , Orthomyxoviridae Infections/immunology , Cytokines/metabolism , RAW 264.7 Cells , Adjuvants, Immunologic/pharmacology , Adjuvants, Immunologic/chemical synthesis , Adjuvants, Immunologic/chemistryABSTRACT
In the development of dendritic cell (DC) vaccines, the maturation of DCs is a critical stage. Adjuvants play a pivotal role in the maturation of DCs, with a major concern being to ensure both efficacy and safety. This study introduces an innovative approach that combines high efficacy with safety through the synthesis of micro-adjuvants grafted with copolymers of 2-(methacrylamido) glucopyranose (MAG) and methacryloxyethyl trimethyl ammonium chloride (DMC). The utilization of metal-free surface-initiated atom transfer radical polymerization enables the production of safe and recyclable adjuvants. These micrometer-sized adjuvants surpass the optimal size range for cellular endocytosis, enabling the retrieval and reuse of them during the ex vivo maturation process, mitigating potential toxicity concerns associated with the endocytosis of non-metabolized nanoparticles. Additionally, the adjuvants exhibit a "micro-ligand-mediated maturation enhancement" effect for DC maturation. This effect is influenced by the shape of the particle, as evidenced by the distinct promotion effects of rod-like and spherical micro-adjuvants with comparable sizes. Furthermore, the porous structure of the adjuvants enables them to function as cargo-carrying "micro-shuttles", releasing antigens upon binding to DCs to facilitate efficient antigen delivery.
Subject(s)
Adjuvants, Immunologic , Dendritic Cells , Polymerization , Dendritic Cells/metabolism , Dendritic Cells/immunology , Adjuvants, Immunologic/chemistry , Adjuvants, Immunologic/pharmacology , Adjuvants, Immunologic/chemical synthesis , Vaccines/chemistry , Vaccines/immunology , Particle Size , Mice , Animals , Polymers/chemistry , Polymers/pharmacology , Polymers/chemical synthesisABSTRACT
Adjuvant is an integral part of all vaccine formulations but only a few adjuvants with limited efficacies or application scopes are available. Thus, developing more robust and diverse adjuvants is necessary. To this end, a new class of adjuvants having α- and ß-rhamnose (Rha) attached to the 1- and 6'-positions of monophosphoryl lipid A (MPLA) was designed, synthesized, and immunologically evaluated in mice. The results indicated a synergistic effect of MPLA and Rha, two immunostimulators that function via interacting with toll-like receptor 4 and recruiting endogenous anti-Rha antibodies, respectively. All the tested MPLA-Rha conjugates exhibited potent adjuvant activities to promote antibody production against both protein and carbohydrate antigens. Overall, MPLA-α-Rha exhibited better activities than MPLA-ß-Rha, and 6'-linked conjugates were slightly better than 1-linked ones. Particularly, MPLA-1-α-Rha and MPLA-6'-α-Rha were the most effective adjuvants in promoting IgG antibody responses against protein antigen keyhole limpet hemocyanin and carbohydrate antigen sTn, respectively.
Subject(s)
Lipid A , Rhamnose , Lipid A/analogs & derivatives , Lipid A/chemistry , Lipid A/pharmacology , Lipid A/immunology , Animals , Rhamnose/chemistry , Rhamnose/immunology , Rhamnose/pharmacology , Mice , Adjuvants, Vaccine/chemistry , Adjuvants, Vaccine/pharmacology , Female , Adjuvants, Immunologic/pharmacology , Adjuvants, Immunologic/chemistry , Adjuvants, Immunologic/chemical synthesis , Toll-Like Receptor 4/metabolism , Toll-Like Receptor 4/agonists , Toll-Like Receptor 4/immunology , Immunoglobulin G/immunology , Immunoglobulin G/blood , Mice, Inbred BALB C , Hemocyanins/chemistry , Hemocyanins/immunologyABSTRACT
Peptide vaccines exhibit great potential in cancer therapy via eliciting antigen-specific host immune response and long-term immune memory to defend cancer cells. However, the low induced immune response of many developing vaccines implies the imperatives for understanding the favorable structural features of efficient cancer vaccines. Herein, we report on the two groups of self-adjuvanting peptide vaccines with distinct morphology and investigate the relationship between the morphology of peptide vaccines and the induced immune response. Two nanofibril peptide vaccines were created via co-assembly of a pentapeptide with a central 4-aminoproline residue, with its derivative functionalized with antigen epitopes derived from human papillomavirus E7 proteins, whereas utilization of a pentapeptide with a natural proline residue led to the formation of two nanoparticle peptide vaccines. The immunological results of dendritic cell (DCs) maturation and antigen presentation induced by the peptide assemblies implied the self-adjuvanting property of the resulting peptide vaccines. In particular, cellular uptake studies revealed the enhanced internalization and elongated retention of the nanofibril peptide vaccines in DCs, leading to their advanced performance in DC maturation, accumulation at lymph nodes, infiltration of cytotoxic T lymphocytes into tumor tissues, and eventually lysis of in vivo tumor cells, compared to the nanoparticle counterparts. The antitumor immune response caused by the nanofibril peptide vaccines was further augmented when simultaneously administrated with anti-PD-1 checkpoint blockades, suggesting the opportunity of the combinatorial immunotherapy by utilizing the nanofibril peptide vaccines. Our findings strongly demonstrate a robust relationship between the immune response of peptide vaccines and their morphology, thereby elucidating the critical role of morphological control in the design of efficient peptide vaccines and providing the guidance for the design of efficient peptide vaccines in the future.
Subject(s)
Adjuvants, Immunologic/pharmacology , Cancer Vaccines/pharmacology , Oropharyngeal Neoplasms/therapy , Papillomaviridae/drug effects , Papillomavirus Infections/therapy , Vaccines, Subunit/pharmacology , Adjuvants, Immunologic/chemical synthesis , Adjuvants, Immunologic/chemistry , Animals , Antigen Presentation/drug effects , Antigen Presentation/immunology , Cancer Vaccines/chemical synthesis , Cancer Vaccines/chemistry , Cell Line , Humans , Immunotherapy , Materials Testing , Mice , Molecular Structure , Neoplasms, Experimental/immunology , Neoplasms, Experimental/therapy , Oropharyngeal Neoplasms/immunology , Papillomaviridae/immunology , Papillomavirus Infections/immunology , Vaccines, Subunit/chemical synthesis , Vaccines, Subunit/chemistryABSTRACT
Modern adjuvants for vaccine formulations are immunostimulating agents whose action is based on the activation of pattern recognition receptors (PRRs) by well-defined ligands to boost innate and adaptive immune responses. Monophosphoryl lipid A (MPLA), a detoxified analogue of lipid A, is a clinically approved adjuvant that stimulates toll-like receptor 4 (TLR4). The synthesis of MPLA poses manufacturing and quality assessment challenges. Bridging this gap, we report here the development and preclinical testing of chemically simplified TLR4 agonists that could sustainably be produced in high purity and on a large scale. Underpinned by computational and biological experiments, we show that synthetic monosaccharide-based molecules (FP compounds) bind to the TLR4/MD-2 dimer with submicromolar affinities stabilizing the active receptor conformation. This results in the activation of MyD88- and TRIF-dependent TLR4 signaling and the NLRP3 inflammasome. FP compounds lack in vivo toxicity and exhibit adjuvant activity by stimulating antibody responses with a potency comparable to MPLA.
Subject(s)
Adjuvants, Immunologic/pharmacology , Glucosamine/pharmacology , Glycolipids/pharmacology , Toll-Like Receptor 4/antagonists & inhibitors , Adaptor Proteins, Vesicular Transport/metabolism , Adjuvants, Immunologic/chemical synthesis , Adjuvants, Immunologic/metabolism , Adjuvants, Immunologic/toxicity , Animals , Female , Glucosamine/chemical synthesis , Glucosamine/metabolism , Glucosamine/toxicity , Glycolipids/chemical synthesis , Glycolipids/metabolism , Glycolipids/toxicity , Humans , Inflammasomes/metabolism , Interleukin-1/metabolism , Macrophages/drug effects , Mice, Inbred C57BL , Myeloid Differentiation Factor 88/metabolism , NLR Family, Pyrin Domain-Containing 3 Protein/metabolism , Signal Transduction/drug effects , Toll-Like Receptor 4/metabolismABSTRACT
Detection of vaccine (adjuvant and antigen) is crucial for the fundamental studies of immunotherapy. In this work, the catechol-containing glycopolymer obtained by sunlight-induced RAFT polymerization was first designed to generate glycoadjuvant@AuNPs. Then, a simple and general self-assembled technique, catechol-driven self-assembly (CDSA), was developed to fabricate AuNP-based glycoadjuvant patterns, regardless of the size, shape and synthetic method of AuNPs. More importantly, highly ordered glycoadjuvant patterns could be easily formed by catechol-driven self-assembly under confinement, which exhibit a higher SERS signal amplification ability for the detection of carbohydrates (glycoadjuvant).
Subject(s)
Adjuvants, Immunologic/chemical synthesis , Catechols/chemistry , Gold/chemistry , Metal Nanoparticles/chemistry , Polymers/chemistry , Adjuvants, Immunologic/chemistry , Molecular Structure , Particle Size , Spectrum Analysis, Raman , Surface PropertiesABSTRACT
In recent years, the utilization of CS-MWCNT as targeted drug carriers has attracted considerable attention. Hericium erinaceus polysaccharide (HEP) has been reported as an immunostimulant to improve immune responses. This study was focussed on developing CS-MWCNT encapsulating HEP (CS-MWCNT-HEP). Using in mice peritoneal macrophages, we found the immune response could be effectively regulated by CS-MWCNT-HEP, promoted the expression of the MHCII, CD86, F4/80 and gp38. Moreover, the mice immunized with CS-MWCNT-HEP nanoparticles significantly extended PCV2-specific IgG immune response and the levels of cytokines. The results demonstrated that CS-MWCNT-HEP may be a promising drug delivery system for immuno-enhancement.
Subject(s)
Adjuvants, Immunologic/chemical synthesis , Fungal Polysaccharides/chemistry , Nanotubes, Carbon/chemistry , Adjuvants, Immunologic/pharmacology , Animals , Antibodies, Viral/immunology , Cells, Cultured , Circovirus/immunology , Cytokines/immunology , Fungal Polysaccharides/immunology , Hericium/chemistry , Immunogenicity, Vaccine , Macrophages, Peritoneal/drug effects , Macrophages, Peritoneal/immunology , Mice , Mice, Inbred ICRABSTRACT
Many studies have investigated how trehalose glycolipid structures can be modified to improve their Macrophage inducible C-type lectin (Mincle)-mediated adjuvanticity. However, in all instances, the ester-linkage of α,ά-trehalose to the lipid of choice remained. We investigated how changing this ester-linkage to an amide influences Mincle signalling and agonist activity and demonstrated that Mincle tolerates this functional group change. In in vivo vaccination studies in murine and ovine model systems, using OVA or Mannheimia haemolytica and Mycoplasma ovipneumoniae as vaccine antigens, respectively, it was demonstrated that a representative trehalose diamide glycolipid was able to enhance antibody-specific immune responses. Notably, IgG titres against M. ovipneumoniae were significantly greater when using trehalose dibehenamide (A-TDB) compared to trehalose dibehenate (TDB). This is particularly important as infection with M. ovipneumoniae predisposes sheep to pneumonia.
Subject(s)
Antibody Specificity/drug effects , Antigens/immunology , Diamide/chemistry , Glycolipids/chemistry , Glycolipids/pharmacology , Lectins, C-Type/metabolism , Membrane Proteins/metabolism , Adjuvants, Immunologic/chemical synthesis , Adjuvants, Immunologic/pharmacology , Animals , Diamide/pharmacology , Gene Expression Regulation/drug effects , Gene Expression Regulation/immunology , Lectins, C-Type/agonists , Lectins, C-Type/genetics , Membrane Proteins/agonists , Membrane Proteins/genetics , Mice , Ovalbumin/immunologyABSTRACT
GM3, a typical tumor-associated carbohydrate antigen, is considered as an important target for cancer vaccine development, but its low immunogenicity limits its application. αGalCer, an iNKT cell agonist, has been employed as an adjuvant via a unique immune mode. Herein, we prepared and investigated two types of antitumor vaccine candidates: (a) self-adjuvanting vaccine GM3-αGalCer by conjugating GM3 with αGalCer and (b) noncovalent vaccine GM3-lipid/αGalCer, in which GM3 is linked with lipid anchor and coassembled with αGalCer. This demonstrated that ßGalCer is an exceptionally optimized lipid anchor, which enables the noncovalent vaccine candidate GM3-ßGalCer/αGalCer to evoke a comparable antibody level to GM3-αGalCer. However, the antibodies induced by GM3-αGalCer are better at recognition B16F10 cancer cells and more effectively activate the complement system. Our study highlights the importance of vaccine constructs utilizing covalent or noncovalent assembly between αGalCer with carbohydrate antigens and choosing an appropriate lipid anchor for use in noncovalent vaccine formulation.
Subject(s)
Adjuvants, Immunologic/pharmacology , Cancer Vaccines/pharmacology , G(M3) Ganglioside/pharmacology , Galactosylceramides/pharmacology , Adjuvants, Immunologic/chemical synthesis , Animals , Cancer Vaccines/chemical synthesis , Cancer Vaccines/immunology , Carbohydrate Sequence , Female , G(M3) Ganglioside/analogs & derivatives , G(M3) Ganglioside/immunology , Galactosylceramides/chemical synthesis , Galactosylceramides/immunology , Humans , Immunity, Humoral/drug effects , Immunoglobulin G/immunology , Liposomes/chemistry , Mice, Inbred BALB C , Natural Killer T-Cells/immunology , THP-1 CellsABSTRACT
Peptide antigens have been widely used in the development of vaccines, especially for those against autoimmunity-inducing pathogens and cancers. However, peptide-based vaccines require adjuvant and/or a delivery system to stimulate desired immune responses. Here, we explored the potential of self-adjuvanting poly(hydrophobic amino acids) (pHAAs) to deliver peptide-based vaccine against Group A Streptococcus (GAS). We designed and synthesized self-assembled nanoparticles with a variety of conjugates bearing a peptide antigen (J8-PADRE) and polymerized hydrophobic amino acids to evaluate the effects of structural arrangement and pHAAs properties on a system's ability to induce humoral immune responses. Immunogenicity of the developed conjugates was also compared to commercially available human adjuvants. We found that a linear conjugate bearing J8-PADRE and 15 copies of leucine induced equally effective, or greater, immune responses than commercial adjuvants. Our fully defined, adjuvant-free, single molecule-based vaccine induced the production of antibodies capable of killing GAS bacteria.
Subject(s)
Adjuvants, Immunologic/therapeutic use , Nanoparticles/therapeutic use , Peptides/therapeutic use , Streptococcal Vaccines/therapeutic use , Streptococcus pyogenes/drug effects , Adjuvants, Immunologic/chemical synthesis , Amino Acid Sequence , Animals , Antigens/therapeutic use , Drug Carriers/chemistry , Drug Carriers/therapeutic use , Female , Immunity, Humoral/drug effects , Immunoglobulin G/immunology , Immunoglobulin G/metabolism , Mice, Inbred C57BL , Nanoparticles/chemistry , Peptides/chemical synthesis , Streptococcal Vaccines/immunology , Streptococcus pyogenes/immunology , Vaccines, SubunitABSTRACT
Mincle agonists have been shown to induce inflammatory cytokine production, such as tumor necrosis factor-alpha (TNF) and promote the development of a Th1/Th17 immune response that might be crucial to development of effective vaccination against pathogens such as Mycobacterium tuberculosis. As an expansion of our previous work, a library of 6,6'-amide and sulfonamide α,α-d-trehalose compounds with various substituents on the aromatic ring was synthesized efficiently in good to excellent yields. These compounds were evaluated for their ability to activate the human C-type lectin receptor Mincle by the induction of cytokines from human peripheral blood mononuclear cells. A preliminary structure-activity relationship (SAR) of these novel trehalose diamides and sulfonamides revealed that aryl amide-linked trehalose compounds demonstrated improved activity and relatively high potency cytokine production compared to the Mincle ligand trehalose dibehenate adjuvant (TDB) and the natural ligand trehalose dimycolate (TDM) inducing dose-dependent and human-Mincle-specific stimulation in a HEK reporter cell line.
Subject(s)
Adjuvants, Immunologic/pharmacology , Lectins, C-Type/agonists , Receptors, Immunologic/agonists , Sulfonamides/pharmacology , Trehalose/analogs & derivatives , Trehalose/pharmacology , Adjuvants, Immunologic/chemical synthesis , HEK293 Cells , Humans , Interleukin-1beta/metabolism , Interleukin-6/metabolism , Small Molecule Libraries/chemical synthesis , Small Molecule Libraries/pharmacology , Sulfonamides/chemical synthesis , Tumor Necrosis Factor-alpha/metabolismABSTRACT
Nucleotide-binding oligomerization domain 2 (NOD2) is cytosolic surveillance receptor of the innate immune system capable of recognizing the bacterial and viral infections. Muramyl dipeptide (MDP) is the minimal immunoreactive unit of murein. NOD2 perceives MDP as pathogen-associated molecular pattern, thereby triggering an immune response with undesirable side-effects. Beneficial properties of MDP, such as pro-inflammatory characteristics for the rational design of new vaccine adjuvants, can be harnessed by strategically re-designing the molecule. In this work, a new class of amphiphilic desmuramylpeptides (DMPs) were synthesized by replacing the carbohydrate moiety (muramic acid) of the parent molecule with hydrophilic arenes. A lipophilic chain was also introduced at the C-terminus of dipeptide moiety (alanine-isoglutamine), while conserving its L-D configuration. These novel DMPs were found to set off the release of higher levels of tumour necrosis factor alpha (TNF-α) than Murabutide, which is a well-known NOD2 agonist. Molecular docking studies indicate that all these DMPs bind well to NOD2 receptor with similar dock scores (binding energy) through a number of hydrogen bonding and hydrophobic/π interactions with several crucial residues of the receptor. More studies are needed to further assess their immunomodulatory therapeutic potential, as well as the possible involvement of NOD2 activation.
Subject(s)
Adjuvants, Immunologic/chemistry , Adjuvants, Immunologic/pharmacology , Peptides/chemistry , Peptides/pharmacology , Adjuvants, Immunologic/chemical synthesis , Humans , Hydrophobic and Hydrophilic Interactions , Molecular Docking Simulation , Nod2 Signaling Adaptor Protein/metabolism , Peptides/chemical synthesis , Surface-Active Agents/chemical synthesis , Surface-Active Agents/chemistry , Surface-Active Agents/pharmacology , THP-1 Cells , Tumor Necrosis Factor-alpha/metabolismABSTRACT
A novel STING agonist, CDGSF, ipsilaterally modified with phosphorothioate and fluorine, was synthesized. The phosphorothioate in CDGSF might be a site for covalent conjugation. Injection of CDGSF generated an immunogenic ("hot") tumor microenvironment to suppress melanoma, more efficiently than dithio CDG. In particular, immunization with SARS-CoV-2 spike protein using CDGSF as an adjuvant elicited an exceptionally high antibody titer and a robust T cell response, overcoming the drawbacks of aluminum hydroxide. These results highlighted the therapeutic potential of CDGSF for cancer immunotherapy and the adjuvant potential of the STING agonist in the SARS-CoV-2 vaccine for the first time.
Subject(s)
Adjuvants, Immunologic/administration & dosage , COVID-19 Vaccines/administration & dosage , COVID-19/prevention & control , Melanoma, Experimental/drug therapy , Membrane Proteins/agonists , Nucleotides, Cyclic/administration & dosage , Skin Neoplasms/drug therapy , Adjuvants, Immunologic/chemical synthesis , Aluminum Hydroxide/administration & dosage , Aluminum Hydroxide/chemistry , Animals , Antibodies, Viral/biosynthesis , B-Lymphocytes/drug effects , B-Lymphocytes/immunology , B-Lymphocytes/virology , COVID-19/immunology , COVID-19/virology , COVID-19 Vaccines/chemistry , Enzyme-Linked Immunospot Assay , Humans , Immunotherapy/methods , Interferon-gamma/biosynthesis , Melanoma, Experimental/immunology , Melanoma, Experimental/mortality , Melanoma, Experimental/pathology , Membrane Proteins/genetics , Membrane Proteins/immunology , Mice , Nucleotides, Cyclic/chemical synthesis , SARS-CoV-2/drug effects , SARS-CoV-2/immunology , SARS-CoV-2/pathogenicity , Skin Neoplasms/immunology , Skin Neoplasms/mortality , Skin Neoplasms/pathology , Spike Glycoprotein, Coronavirus/administration & dosage , Spike Glycoprotein, Coronavirus/chemistry , Spike Glycoprotein, Coronavirus/immunology , Survival Analysis , T-Lymphocytes/drug effects , T-Lymphocytes/immunology , T-Lymphocytes/virology , Tumor Burden/drug effects , Tumor Microenvironment/drug effects , Tumor Microenvironment/immunology , Vaccination/methodsABSTRACT
The COVID-19 pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has led to an unprecedented effort toward the development of an effective and safe vaccine. Aided by extensive research efforts into characterizing and developing countermeasures towards prior coronavirus epidemics, as well as recent developments of diverse vaccine platform technologies, hundreds of vaccine candidates using dozens of delivery vehicles and routes have been proposed and evaluated preclinically. A high demand coupled with massive effort from researchers has led to the advancement of at least 31 candidate vaccines in clinical trials, many using platforms that have never before been approved for use in humans. This review will address the approach and requirements for a successful vaccine against SARS-CoV-2, the background of the myriad of vaccine platforms currently in clinical trials for COVID-19 prevention, and a summary of the present results of those trials. It concludes with a perspective on formulation problems which remain to be addressed in COVID-19 vaccine development and antigens or adjuvants which may be worth further investigation.
Subject(s)
Adjuvants, Immunologic/chemical synthesis , COVID-19 Vaccines/chemical synthesis , COVID-19/prevention & control , Drug Development/methods , SARS-CoV-2/drug effects , Adjuvants, Immunologic/therapeutic use , Animals , COVID-19/immunology , COVID-19 Vaccines/therapeutic use , Drug Compounding/methods , Drug Compounding/trends , Drug Development/trends , Humans , Recombinant Proteins/chemical synthesis , Recombinant Proteins/therapeutic use , SARS-CoV-2/immunologyABSTRACT
Adjuvants play a critical role in the design and development of novel vaccines. Despite extensive research, only a handful of vaccine adjuvants have been approved for human use. Currently used adjuvants are mostly composed of components that are non-native to the human body, such as aluminum salt, bacterial lipids, or foreign genomic material. Here, a new ionic-liquid-based adjuvant is explored, synthesized using two metabolites of the body, choline and lactic acid (ChoLa). ChoLa distributes the antigen efficiently upon injection, maintains antigen integrity, enhances immune infiltration at the injection site, and leads to a potent immune response against the antigen. Thus, it can serve as a promising safe adjuvant platform that can help to protect against pandemics and future infectious threats.
Subject(s)
Adjuvants, Immunologic/chemistry , Ionic Liquids/chemistry , Safety , Adjuvants, Immunologic/chemical synthesis , Animals , Glycerylphosphorylcholine/chemistry , Lactic Acid/chemistryABSTRACT
In aqueous conditions, amphiphilic bioactive molecules are able to form self-assembled colloidal structures modifying their biological activity. This behavior is generally neglected in preclinical studies, despite its impact on pharmacological development. In this regard, a significative example is represented by a new class of amphiphilic marine-inspired vaccine adjuvants, collectively named Sulfavants, based on the ß-sulfoquinovosyl-diacylglyceride skeleton. The family includes the lead product Sulfavant A (1) and two epimers, Sulfavant R (2) and Sulfavant S (3), differing only for the stereochemistry at C-2 of glycerol. The three compounds showed a significant difference in immunological potency, presumably correlated with change of the aggregates in water. Here, a new synthesis of diastereopure 3 was achieved, and the study of the immunomodulatory behavior of mixtures of 2/3 proved that the bizarre in vitro response to 1-3 effectively depends on the supramolecular aggregation states, likely affecting the bioavailability of agonists that can effectively interact with the cellular targets. The evidence obtained with the mixture of pure Sulfavant R (2) and Sulfavant S (3) proves, for the first time, that supramolecular organization of a mixture of active epimers in aqueous solution can bias evaluation of their biological and pharmacological potential.
Subject(s)
Adaptive Immunity/drug effects , Adjuvants, Immunologic/pharmacology , Dendritic Cells/drug effects , Adjuvants, Immunologic/chemical synthesis , Biomarkers/metabolism , Cells, Cultured , Colloids , Dendritic Cells/immunology , Dendritic Cells/metabolism , Humans , Molecular Structure , Phenotype , Solubility , Structure-Activity RelationshipABSTRACT
Adjuvants are a crucial component of recombinant vaccines such as the human papillomavirus (HPV) vaccine. Monophosphoryl lipid A (MPL) extracted from Salmonella Minnesota lipopolysaccharide is used as an adjuvant for the HPV vaccine. Due to the limitations in accessibility and reproducibility of MPL, investigating synthetic analogues of MPL (synMPL) is urgently needed to overcome these limitations. In this study, female BALB/c mice were vaccinated by HPV vaccine formulated with synMPL and aluminum hydroxide gel in which the concentration of synMPL ranged from 0 to 100 µg/dose. Anti-HPV L1 VLP antibody was measured for each group through Indirect ELISA and compared with Cervarix and Gardasil vaccines as approved anti-HPV vaccines. SynMPL showed a concentration-dependent increase up to 50 µg/dose in the immunogenicity of the vaccine. Therefore, synMPL at concentration of 50 µg/dose was selected as optimum concentration. The GMT profiling of synMPL-formulated vaccine (named Papilloguard) and Cervarix was not statistically different (Mann-Whitney test). The Gardasil vaccine showed 10-fold lower GMT for anti-HPV 18 L1 VLP antibody but anti-HPV 16 L1 VLP antibody was similar to Cervarix and Papilloguard. The current findings suggest that the synMPL in combination with aluminum hydroxide could be used as a potential adjuvant candidate for human vaccine.