CN117244054A

CN117244054A - Adjuvant use of compounds having spirostane-O-gal structure

Info

Publication number: CN117244054A
Application number: CN202311336011.2A
Authority: CN
Inventors: 王志标; 李月; 王欣瑀; 王旭; 单璞; 陈金燕; 李树香
Original assignee: China National Biotec Research Institute Co ltd
Current assignee: China National Biotec Research Institute Co ltd
Priority date: 2023-10-16
Filing date: 2023-10-16
Publication date: 2023-12-19

Abstract

The invention discloses application of a compound or a pharmaceutically acceptable derivative thereof in preparing a nonspecific immune response enhancer. The compound shown as the formula I in the disclosure is used as an adjuvant, has milder action compared with the marketed adjuvant, and does not induce serious inflammatory reaction at the injection site. The overall effect is not lower than that of an aluminum adjuvant, but compared with the defect that the aluminum adjuvant is easy to accumulate and is not easy to metabolize, the small molecular compound shown in the formula I is easier to metabolize, and the safety is better. Compared with the traditional aluminum adjuvant, the compound shown in the formula I has natural hydrophilic and hydrophobic groups, is easy to penetrate through biological barriers, and has wider application range and higher application value as the adjuvant.

Description

Adjuvant use of compounds having spirostane-O-gal structure

Technical Field

The present disclosure relates to the field of pharmaceutical technology, in particular, the disclosure relates to the use of multiple compounds having the same structural unit as non-specific immune response enhancers.

Background

Vaccine immunization is one of the main tools for reducing pathogen transmission and improving the health level of people. However, the immune system is affected by the inoculation mode, the times, the immune stress and other factors, and the situation that the immune organs of the organism are damaged and the effective immune protection cannot be achieved after the inoculation possibly occurs in the immune process. Vaccine adjuvants are an effective means to solve the above problems. The adjuvant is an immunostimulant for enhancing and regulating the immune response of vaccine antigens, and can generate stronger immune response when used together with the antigens, so that the dosage of the antigens can be reduced, the times of vaccination can be reduced, and the safety of the vaccine can be improved. In addition to traditional aluminum adjuvants, novel adjuvants such as oil-in-water emulsions, lipids, saponins, nucleic acids, etc. are being continuously developed and used. However, although a variety of vaccine adjuvants have been used clinically, they suffer from a variety of drawbacks in different respects. For example, clinically common aluminum adjuvants produce weaker immune responses and cannot participate in cellular immune responses than other applied adjuvants; studies have revealed that aluminum adjuvants can enter the brain in white mice, with risks. The type of the induced immune reaction is biased (Th 2 type), the local reaction is heavy, and meanwhile, the aluminum adjuvant is easy to form granuloma, even local aseptic abscess can occur, and the allergic reaction of the organism is caused.

The saponin is a glycoside compound extracted from natural plants, is an active ingredient of a plurality of Chinese herbal medicines such as ginseng, bitter orange, platycodon root, liquorice and the like, and has various medicinal functions such as anti-inflammatory, anti-tumor, liver and kidney protection, immunoregulation and the like. The number of saponins currently found is up to several tens of thousands. In terms of vaccine immunization, QS-21 saponin isolated from the bark of quillaja saponaria has been found to have significant immune activity, eliciting strong humoral and T cell responses, and thus has been approved for malaria vaccines and herpes zoster vaccines, suggesting that such substances have great potential as vaccine adjuvants. Compared with the traditional aluminum adjuvant, the saponin substance is easy to penetrate through biological barriers due to natural hydrophilic and hydrophobic groups, and has wider application as an adjuvant (such as an in-situ vaccine adjuvant for solid tumors). However, QS-21 adjuvants also have certain drawbacks in clinical use, such as the susceptibility to severe inflammatory reactions at the site of injection, toxicity and poor hemolysis, and difficulty in production and synthesis due to a single source. Therefore, finding a new substance that can replace the existing known adjuvants has become one of the important targets for adjuvant development.

Disclosure of Invention

The technical problems to be solved are as follows:

in one aspect of the present disclosure, the present disclosure provides a novel adjuvant use of compounds, which is directed to the disadvantage of weak immune response and susceptibility to inflammatory reaction of prior art adjuvants such as aluminum adjuvants or saponin-derived QS-21.

In particular, the present inventors have surprisingly found in the study that among tens of thousands of saponins, a series of saponins having a parent nucleus as shown in formula I and derivatives thereof undergo only slight inflammatory cell recruitment after injection, and are capable of inducing an antigen to generate excellent immune response in the body, and have good neutralizing antibody titer. The discovery of the application of the series of compounds as nonspecific immune response enhancers solves the defects of the adjuvants in the prior art, and is hopeful to be developed into a novel vaccine adjuvant.

The technical scheme is as follows:

the use of a compound or a pharmaceutically acceptable derivative thereof for the preparation of a non-specific immune response enhancer, said compound being of formula I,

wherein,

R ₁ independently selected from H, a substitutable aryl or heteroaryl group;

R ₂ independently selected from H, a substitutable aryl or heteroaryl group;

R ₃ independently selected from H, a substitutable aryl or heteroaryl group;

R ₄ is H or OH;

R ₅ h or OH.

In the present disclosure, the compound represented by formula I or a pharmaceutically acceptable derivative thereof is a saponin compound.

Preferably, in certain embodiments of the present disclosure:

the R is ₁ Is independently selected from H,

The R is ₂ Is independently selected from H,

The R is ₃ Independently selected from H, OH or COH.

More preferably, in certain embodiments of the present disclosure:

the compound is selected from the following compounds:

further preferably, in certain embodiments of the present disclosure:

the compound is selected from the following compounds:

in the present disclosure, the pharmaceutically acceptable derivative may be in the form of a pharmaceutically acceptable acid or basic salt thereof. Preferably, in certain embodiments of the present disclosure, the acid salt is a hydrochloride, sulfate, phosphate, citrate, hydrobromide, acetate, benzoate, benzenesulfonate, tartrate, carbonate, citrate, gluconate, lactate, malate, methanesulfonate, stearate, valerate, or nitrate; the basic salt is sodium salt, calcium salt, potassium salt, zinc salt or meglumine salt.

In the present disclosure, the compounds may also include forms of isomers, hydrates, solvates, metabolites, or prodrugs thereof.

In another aspect of the present disclosure, there is provided the use of a compound as described above, or a pharmaceutically acceptable derivative thereof, in the preparation of an immunogenic composition or pharmaceutical composition.

In certain embodiments of the present disclosure, the immunogenic or pharmaceutical composition may be a composition comprising an inactivated vaccine, an attenuated live vaccine, a protein vaccine, a bacterial polysaccharide and polysaccharide protein conjugate vaccine, a genetically engineered vaccine, or a genetically reassortant vaccine.

In another aspect of the present disclosure, there is provided a vaccine composition comprising an immunogenic material and a compound as described above or a pharmaceutically acceptable derivative thereof.

In another aspect of the present disclosure, there is provided a compound adjuvant comprising the above compound or a pharmaceutically acceptable derivative thereof, and other non-specific immune response enhancers.

Preferably, in certain embodiments of the present disclosure, the other non-specific immune response enhancer is one or more selected from the group consisting of aluminum adjuvants, calcium adjuvants, liposomes, MPL, MF-59, SAF, freund's adjuvants, saponin adjuvants, manganese salt adjuvants, AS03, AS04, tween 80, bacterial lipopolysaccharide adjuvants, RIBI adjuvant systems, cpG-ODN adjuvants, imidazopyridine compounds.

In another aspect of the present disclosure, there is provided a method of inducing an immune response in a mammal, comprising administering to the mammal an effective amount of an immunogenic composition, pharmaceutical composition or vaccine composition as described above.

The beneficial effects are that:

the compound shown as the formula I in the disclosure is used as an adjuvant, has milder action compared with the marketed adjuvant, and does not induce serious inflammatory reaction at the injection site. The overall effect is not lower than that of an aluminum adjuvant, but compared with the defect that the aluminum adjuvant is easy to accumulate and is not easy to metabolize, the small molecular compound shown in the formula I is easier to metabolize, and the safety is better. Compared with the traditional aluminum adjuvant, the compound shown in the formula I has natural hydrophilic and hydrophobic groups, is easy to penetrate through biological barriers, and has wider application range and higher application value as the adjuvant.

Drawings

FIG. 1 is a graph showing the results of inflammatory response after intramuscular injection of various samples in example 2 of the present disclosure;

FIG. 2 is a graph showing the results of inflammatory response after intramuscular injection of various samples in example 7 of the present disclosure;

FIG. 3 is a graph comparing the results of neutralizing antibodies generated after the immunization of different compounds against HBsAg antigen in the examples of the present disclosure;

FIG. 4 is a graph comparing the results of neutralizing antibodies generated after the immunization of different compounds against the Covid-19S-trimer antigen in the examples of the present disclosure;

FIG. 5 is a graph showing the results of inflammatory responses after intramuscular injection of different samples in the examples of the present disclosure;

FIG. 6 is a graph comparing the results of neutralizing antibodies generated after the immunization of different compounds against HBsAg antigen in the examples of the present disclosure;

FIG. 7 is a graph comparing the results of the neutralizing antibodies generated after the immunization of different compounds against the H1N1 antigen in the examples of the present disclosure;

FIG. 8 is a graph comparing the results of neutralizing antibodies generated after the immunization of various compounds against the Covid-19S-trimer antigen in the examples of the present disclosure.

Detailed Description

The invention discloses application of a compound or a pharmaceutically acceptable derivative thereof in preparing a nonspecific immune response enhancer, and a person skilled in the art can properly improve process parameters by referring to the content of the compound or the pharmaceutically acceptable derivative. It is to be particularly pointed out that all similar substitutes and modifications apparent to those skilled in the art are deemed to be included in the invention and that the relevant person can make modifications and appropriate alterations and combinations of what is described herein to make and use the technology without departing from the spirit and scope of the invention.

In the present invention, unless otherwise indicated, scientific and technical terms used herein have the meanings commonly understood by one of ordinary skill in the art. Throughout the specification and claims, unless explicitly stated otherwise, the term "comprise" or variations thereof such as "comprises" or "comprising", etc. will be understood to include the stated element or component without excluding other elements or components. The terms "such as," "for example," and the like are intended to refer to exemplary embodiments and are not intended to limit the scope of the present disclosure.

Definition:

the terms "adjuvant", "vaccine adjuvant", "immunoadjuvant" in this disclosure refer to a class of substances that are capable of non-specifically binding or mixing with an antigen/immunogen/immunogenic substance, enhancing the immunogenicity and immunoprotection effects of the antigen/immunogen/immunogenic substance, i.e. enhancing humoral and/or cellular immune responses, while being self-non-immunogenic. It is also understood as "substances which are used to bind to an antigen and which give better immunity than the antigen alone".

In the present disclosure, the subjects to which the "adjuvant" (adjvant), "vaccine adjuvant", "immunoadjuvant", nonspecific immune response enhancer, etc., are administered are any mammals including, but not limited to, domestic and farm animals (cattle, horses, pigs, sheep, goats, dogs, cats, rodents, etc.), primates, and humans (homosapiens). Preferably, the mammal is a human.

The term "aryl" refers to a single all-carbon aromatic ring or a polycyclic fused all-carbon ring system in which at least one of the rings is aromatic. For example, in certain embodiments of the present disclosure, the aryl group has 6 to 20 carbon atoms, 6 to 14 carbon atoms, or 6 to 12 carbon atoms. In one embodiment of the present disclosure, aryl includes phenyl. In other embodiments of the present disclosure, aryl groups further include polycyclic fused ring systems having about 9 to 20 carbon atoms (e.g., ring systems comprising 2,3, or 4 rings), wherein at least one ring is aromatic, and wherein the other rings may or may not be aromatic (i.e., carbocyclic).

The term "heteroaryl" refers to a single aromatic ring having at least one atom other than carbon in the ring, wherein the atoms are selected from the group consisting of oxygen, nitrogen, and sulfur. In certain embodiments, non-limiting examples of heteroaryl groups include: thienyl (thiophen); benzo [ b ] thienyl; naphtho [2,3-b ] thienyl; a thianthrene group; furyl (furanyl); isobenzofuranyl; a chromene group; xanthenyl; phenoxathia group; pyrrolyl, including but not limited to 2H-pyrrolyl; imidazolyl; pyrazolyl; pyridyl (pyridyl, pyridinyl) including, but not limited to, 2-pyridyl, 3-pyridyl and 4-pyridyl; pyrazinyl; pyrimidinyl; a pyridazinyl group; indolizinyl; isoindolyl; 3H-indolyl; indolyl; indazolyl; a purine group; 4H-quinolizinyl; isoquinolinyl; quinolinyl; 2, 3-naphthyridinyl; 1, 5-naphthyridinyl; quinazolinyl; cinnolinyl; pteridinyl; carbazolyl; beta-carboline groups; phenanthridinyl; an acridine group; a naphthyridine group; phenanthroline group; a phenazinyl group; isothiazolyl; a phenothiazinyl group; isoxazolyl; furazanyl; a phenoxazinyl group; 1, 4-dihydroquinoxaline-2, 3-dione; 7 amino isocoumarins; pyrido [1,2-a ] pyrimidin-4-one; pyrazolo [1,5-a ] pyrimidinyl, including but not limited to pyrazolo [1,5-a ] pyrimidin-3-yl; 1, 2-benzisoxazol-3-yl; benzimidazolyl; 2-oxindolyl and 2-oxo-benzimidazolyl.

The term "substitutable" means that a hydrogen group of a specified moiety may be replaced by a group of a specified substituent, provided that the substitution results in a stable or chemically feasible compound.

The term "acyl" refers to the radical remaining after removal of one or more hydroxyl groups of an organic or inorganic oxy acid of the formula R-M (O) -. In certain embodiments, non-limiting examples of "acyl" include acetyl, propionyl, butyryl, isobutyryl, benzoyl, 1-naphthoyl, 2-naphthoyl, and the like.

The term "aralkyl" refers to any C1-10 alkyl group substituted with any C6-14 aryl group. In certain embodiments, non-limiting examples of aralkyl groups include benzyl, phenethyl, and naphthylmethyl.

The term "aliphatic" includes saturated and unsaturated straight-chain (i.e., unbranched), branched, acyclic, cyclic, or polycyclic aliphatic hydrocarbons, optionally substituted with one or more functional groups. In certain embodiments, "aliphatic" is intended herein to include, but is not limited to, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, and cycloalkynyl, as will be appreciated by those of ordinary skill in the art.

The term "cycloaliphatic radical" refers to a cyclic, non-aromatic, saturated or unsaturated hydrocarbon radical having typically 3 to 20 ring carbon atoms. In certain embodiments, non-limiting examples include cycloalkanes, cycloalkenes, and cycloalkynes. The cycloaliphatic radical may also comprise heteroatoms or heteroatom groups selected from N, O, S and SO 2.

The term "heterocyclyl" refers to groups having 3-to 10-membered non-aromatic ring systems of ring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, sulfur, boron, phosphorus and silicon.

The term "glycosyl" refers to a monovalent substituent formed by removing a hemiacetal hydroxyl group from a cyclic form of a monosaccharide (or disaccharide, trisaccharide), such as a glycosyl to replace one or more OH groups on a compound of formula I. In certain embodiments, non-limiting examples include pentosyl, hexoyl, beta-D glucopyranosyl.

The term "isomer" refers to compounds of the same chemical formula having the same chemical bonds but having different arrangements of atoms. Isomers can be divided into (carbon) chain isomerism, positional isomerism and functional isomerism.

The term "hydrate" refers to compounds of the present disclosure and pharmaceutically acceptable derivatives thereof also include stoichiometric or non-stoichiometric amounts of water bound by non-covalent intermolecular forces.

The term "solvate" refers to an association of one or more solvent molecules with a compound of the present disclosure and pharmaceutically acceptable derivatives thereof. In certain embodiments, the solvent that forms the solvate includes, but is not limited to, water, isopropanol, ethanol, methanol, dimethyl sulfoxide, ethyl acetate, acetic acid, aminoethanol.

The term "metabolite" refers to the product of the compounds of the present disclosure and pharmaceutically acceptable derivatives thereof, obtained in vivo by metabolism.

The term "prodrug" also refers to a prodrug, a prodrug and the like, and refers to a compound which is obtained by chemical structure modification of a drug, is inactive or less active in vitro, and releases an active drug in vivo through enzymatic or non-enzymatic conversion to exert a drug effect.

Immunogenic composition:

in the present disclosure, the "immunogenic composition" includes "vaccine composition" that is capable of eliciting, activating, and enhancing an immune response in the body. The immune response includes a cellular immune response and/or a humoral immune response. The immune response may be a prophylactic immune response or a therapeutic immune response.

In the present disclosure, the "immunogenic composition" includes an immunogenic substance, and a compound represented by formula 1 or a pharmaceutically acceptable derivative thereof as a nonspecific immune response enhancer. In certain embodiments of the present disclosure, non-limiting examples of the immunogenic material include natural, recombinant, or synthetic products derived from viruses, bacteria, fungi, and parasites, and fragments or portions thereof. In other embodiments of the present disclosure, the immunogenic material may be a peptide, peptide/MHC molecule complex. In other embodiments of the present disclosure, the immunogenic material may be in a particulate form, such as a virus-like particle (VLP).

When the immunogenic agent is a virus, in certain embodiments of the present disclosure, non-limiting examples of immunogenic agents include any of hepatitis a, hepatitis B, hepatitis c, influenza, varicella, adenovirus, herpes simplex virus type I (HSV I), herpes simplex type II (HSV II), vaccinia, rhinovirus, epstein-barr virus, rotavirus, respiratory syncytial virus, papilloma virus, papova virus, cytomegalovirus, arbovirus, hantavirus, coxsackie virus, mumps virus, measles virus, rubella virus, polio virus, human immunodeficiency virus type I (HIV I) and human immunodeficiency virus type II (HIV II), any of picornaviridae, enterovirus, norwalk virus, togavirus, such as alphavirus, flaviviridae, coronavirus, rabies virus, marburg virus, ebola virus, paramyxovirus, bunyavirus, arenavirus, reovirus, rotavirus, leukemia virus, human T cell type I, human immunodeficiency virus, simian virus, herpes virus 1, and herpes virus (epstein virus).

In the present disclosure, the ratio of the immunogenic material in the "immunogenic composition" to the compound represented by formula 1 or a pharmaceutically acceptable derivative thereof may be 1:1 to 100 (wt). In certain embodiments of the present disclosure, the ratio may be 1:5, 1:10, 1:15, 1:20, 1:25, 1:30, 1:35, 1:40, 1:45, 1:50, 1:55, 1:60, 1:65, 1:70, 1:75, 1:80, 1:85, 1:90, 1:95, 1:100 (wt). The administration thereof may be, for example, oral, intravenous, arterial, mucosal, nasal, intramuscular, subcutaneous, organ or intraperitoneally in certain embodiments of the invention. The immunogenic composition may be formulated according to conventional methods into any suitable dosage form, as desired.

In the present disclosure, the administered dose of the "immunogenic composition" should be titrated according to the actual situation. In general, for example, in certain embodiments of the invention, the dosing may range from 0.00001mg to 100mg per kg of subject body weight. The frequency of administration may be, for example, in certain embodiments of the invention, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 8 months, 10 months, 1 year or longer. Often, the blood concentration of the immunogenic composition and/or the memory of the immune cells may be increased by multiple administrations.

Composite adjuvant:

a complex adjuvant is typically a complex formed by two or more adjuvants of the same or different mechanisms to better elicit, activate, and enhance an immune response in the body. In the present disclosure, the compound adjuvant includes a compound represented by formula 1 as set forth in claim 1 or a pharmaceutically acceptable derivative thereof, as well as other adjuvants.

Saponins:

saponins (saponines), a class of glycosides whose aglycones are triterpenes or spirostanes, consist of hydrophilic regions (usually several sugar chains) bound to hydrophobic regions of the steroid or triterpene structure. The most major saponins used in the art for the manufacture of vaccines are those derived from the plants southern American tree Quillaja saponaria (Quillaja saponaria) (Molina), aesculus hippocastanum (Aesculus hippocastanum) or Gyophilla struthium. In the present disclosure, the compounds of formula I are a class of sapogenins having hederagenin-O-Ara parent cores, which have been found by the present inventors to have excellent immunoadjuvant functions. In addition to the compounds of formula I, in certain embodiments of the present disclosure, there is provided the use of several compounds similar in structure thereto for the preparation of a non-specific immune response enhancer, comprising:

in still other embodiments of the present disclosure, there is also provided the use of several other compounds for the preparation of a non-specific immune response enhancer, comprising:

in order to enable those skilled in the art to better understand the technical solution of the present invention, the present invention will be further described in detail with reference to specific embodiments.

Examples:

this example demonstrates the properties of the compounds shown in table 1 below within the scope of the claims.

Compounds of table 1

Example 1: preparation of immunogenic compositions

The following immunogenic compositions were prepared separately, and the compounds were dissolved with a suitable solvent, for example, water, DMSO, 0.9% NaCl solution, and the like. And uniformly mixing the dissolved compound with the antigen.

1. Hepatitis b virus recombinant surface antigen (HBsAg) immunogenic composition: see table 2.

TABLE 2

Grouping	HBsAg dose	Dosage of the compound
			Compound (1-4) +HBsAg	0.5μg	5μg
QS-21+HBsAg	0.5μg	5μg
			Alhydrogel+HBsAg	0.5μg	25μg(Al)
0.9％NaCl+HBsAg	0.5μg	0μg

Hepatitis B virus recombinant surface antigen (HBsAg) source Beijing biological product research all responsible companies; aluminum adjuvant Alhydrogel was from Croda corporation.

2. H1N1 influenza virus lytic subunit antigen (H1N 1) immunogenic composition: see table 3.

TABLE 3 Table 3

Grouping	H1N1 dose	Dosage of the compound
			Compounds (1-4) +H2N 1	0.3μg	5μg
Alhydrogel+H1N1	0.3μg	25μg(Al)
			0.9％NaCl+H1N1	0.3μg	0μg

H1N1 influenza virus lytic subunit antigen (H1N 1) source vinca biologicals research all responsible companies; aluminum adjuvant Alhydrogel was from Croda corporation.

3. Novel coronavirus Omicron strain recombinant S-trimer (S-trimer) immunogenic composition: see table 4.

TABLE 4 Table 4

Novel coronavirus Omicron strain recombinant S-trimer (S-trimer) source beijing Yiqiao shenzhou science and technology company limited; aluminum adjuvant Alhydrogel was from Croda corporation.

Example 2: recruitment of compounds to inflammatory cells at the site of injection

Samples of compound 2, compound 4, and QS-21 (MCE) and 0.9% NaCl are taken as examples. After dissolving the samples, 5 μg each was taken and injected intramuscularly on the outside of the rat thigh, after 72 hours, the leg muscles were separated, sectioned after fixation, and the change in muscle tissue was observed under the HE-stained mirror. QS-21 group served as positive control and physiological saline group served as negative control. The results are shown in FIG. 1. As a result, it was found that the QS-21 injection group showed shrinkage of muscle fibers and massive inflammatory cell infiltration, compared to the negative control group. Whereas the compounds disclosed in this disclosure only undergo slight inflammatory cell recruitment, indicating that they would be more safe than QS-21.

Example 3: enhancement of immune response of compounds to chicken Ovalbumin (OVA) immunogens

Balb/c mice (Beijing vernonihua laboratory animal technologies Co., ltd.) of 6-8 weeks old were randomly grouped, 7 mice per group were immunized by thigh intramuscular injection at 0 week, 3 weeks, and blood was collected 1 week after the last immunization, and the IgG antibody titer in serum was measured by ELISA. The immunization doses are shown in table 5. Chicken Ovalbumin (OVA) was from invitrogen.

TABLE 5

Grouping	Antigen dose	Adjuvant dosage
			Al adjuvant+OVA group	25 μg/mouse	50 μg (aluminium content)/mouse
Compound adjuvant + OVA group	25 μg/mouse	10 μg (compound)/mouse

The results are shown in Table 6. The results showed that the Al adjuvant group antibody titres were 201ng/ml and that 10. Mu.g of each compound adjuvant group induced antibody titres greater than 50. Mu.g of aluminum adjuvant.

TABLE 6

Immunogenic compositions	OVA immunocompetence (IgG ng/ml)
		Compound 1+OVA	261
Compound 2+OVA	280
		Compound 3+OVA	542
Compound 4+OVA	840
		Al+OVA	201

Example 4: immune potentiation of recombinant hepatitis B virus surface antigen (HBsAg) by Compounds

An immunogenic composition was prepared according to (1) of example 1. Babl/c mice with the age of 6-8 weeks are randomly divided into 6 groups of 5 mice. Two immunizations (5. Mu.g of compound, 0.5. Mu.g of antigen, 25. Mu.g of aluminum adjuvant) were performed at weeks 0 and 3, blood was collected at week 5, and then IgG antibody titer was determined.

The results are shown in Table 7. Analysis shows that the antibody can induce strong antibody titer, the average titer is improved by about 100 times compared with the antigen group alone, and the average titer is equivalent to or better than that of 25 mug of aluminum adjuvant.

TABLE 7

Immunogenic compositions	Log anti-HBs(mIU/ml)
		Compound 1+HBsAg	2.45
Compound 2+HBsAg	3.32
		Compound 3+HBsAg	3.02
Compound 4+HBsAg	3.04
		QS-21+HBsAg	3.66
Alhydrogel+HBsAg	2.87
		0.9％NaCl+HBsAg	1.37

Example 5: immune potentiation of compounds against H1N1 influenza virus lytic subunit antigen (H1N 1)

An immunogenic composition was prepared according to (2) of example 1. Babl/c mice with the age of 6-8 weeks are randomly divided into 5 groups of 5 mice each. Two immunizations (5. Mu.g of compound, 0.3. Mu.g of antigen, 25. Mu.g of aluminum adjuvant) were performed at weeks 0 and 3, blood was collected at week 5, and then blood-inhibitory titers were determined using fresh chicken blood.

The results are shown in Table 8. The results show that a fraction of 5 μg of compound as adjuvant induced a neutralizing antibody titer higher than 25 μg of aluminum adjuvant, and both far higher than the antigen group alone.

TABLE 8

Immunogenic compositions	Neutralizing antibody titre
		Compound 1+H1N1	11.2
Compound 2+H1N1	89.6
		Compound 3+H1N1	23.2
Compound 4+H1N1	118.4
		Alhydrogel+H1N1	57.6
0.9％NaCl+H1N1	7.6

Example 6: immune potentiation of novel coronavirus Omicron strain recombinant S-trimer (S-trimer) by compounds Action

An immunogenic composition was prepared according to (3) of example 1. Babl/c mice with the age of 6-8 weeks are randomly divided into 6 groups of 5 mice. Two immunizations (5. Mu.g of compound, 0.3. Mu.g of antigen, 25. Mu.g of aluminum adjuvant) were performed at weeks 0 and 2, blood was collected at week 3, and then IgG antibody titer was determined.

The results are shown in Table 9. The results show that the compound has good promoting effect on the immune effect of the antigen, and compared with the single antigen group, the antibody titer can be obviously improved.

TABLE 9

Immunogenic compositions	Lg endpoint titers
		Compound 1+S-trimer	3.2
Compound 2+S-trimer	4.4
		Compound 3+S-trimer	4.0
Compound 4+S-trimer	3.8
		Alhydrogel+S-trimer	5.4
0.9％NaCl+S-trimer	2.2

Example 7: evaluation of adjuvant function of Picria Sonchifolia saponin B and ginsenoside Rg5

1. Recruitment of inflammatory cells to injection sites

Picria fel-terrae saponin B, ginsenoside Rg5 samples, QS-21 (MCE company) and 0.9% NaCl were taken. After dissolving the samples, 5 μg each was taken and injected into the thigh outside muscle of the same batch of mice, after 72 hours, the leg muscles were separated, sectioned after fixation, and the change in muscle tissue was observed under HE staining and a scope. QS-21 group served as positive control and physiological saline group served as negative control. The results are shown in FIG. 2. As a result, it was found that the QS-21 injection group showed shrinkage of muscle fibers and massive inflammatory cell infiltration, compared to the negative control group. Whereas the compounds disclosed in this disclosure only undergo slight inflammatory cell recruitment, indicating that they would be more safe than QS-21.

2. Immune potentiation of recombinant hepatitis B virus surface antigen (HBsAg)

The following immunogenic compositions were prepared according to Table 10, and the compounds were dissolved with appropriate solvents, e.g., water, DMSO, 0.9% NaCl solution, etc. And uniformly mixing the dissolved compound with the antigen.

Table 10

The same batch of Babl/c mice with the age of 6-8 weeks are randomly divided into 5 groups of 5 mice each. Gold standard Alhydrogel using aluminum adjuvant and above was saponin adjuvant as positive control, and was immunized twice at 0 week and 3 weeks, blood was collected at 5 weeks, and then antigen-specific IgG antibody titer was measured. The results of antibody titres are shown in FIG. 3, and it is found by analysis that the above compounds can induce strong antibody titres when mixed with HBsAg. The average antibody titer was slightly higher than that of aluminum adjuvant, and there was no significant difference with QS-21, demonstrating the excellent adjuvant effect of the above compounds.

3. Immune potentiation of novel coronavirus Omicron strain recombinant S-trimer

The following immunogenic compositions were prepared according to Table 11, and the compounds were dissolved with appropriate solvents, e.g., water, DMSO, 0.9% NaCl solution, etc. And uniformly mixing the dissolved compound with the antigen.

TABLE 11

The same batch of Babl/c mice with the age of 6-8 weeks is taken and randomly divided into 6 groups of 5 mice. Two immunizations (5. Mu.g of compound, 0.3. Mu.g of antigen, 25. Mu.g of aluminum adjuvant) were performed at weeks 0 and 2, blood was collected at week 3, and then IgG antibody titer was determined. The analysis results are shown in fig. 4, and the analysis shows that when the novel coronavirus recombinant S trimer antigen is matched, compared with a single antigen group, 5 mug picriafel-terrae glycoside IB can obviously improve the antibody titer (the average value is improved by more than 100 times), and compared with 25 mug aluminum adjuvant, the antibody titer is not obviously different.

Example 8: adjuvant function evaluation of Polygalasaponin D, saikosaponin B2, saikosaponin E, aescin Ie

1. Recruitment of inflammatory cells to injection sites

A sample of polygalasaponin D, saikosaponin B2, saikosaponin E, aescine Ie, QS-21 (MCE Co.) and 0.9% NaCl was taken. After dissolving the samples, 5 μg each was taken and injected into the thigh outside muscle of the same batch of mice, after 72 hours, the leg muscles were separated, sectioned after fixation, and the change in muscle tissue was observed under HE staining and a scope. QS-21 group served as positive control and physiological saline group served as negative control. The results are shown in FIG. 5. As a result, it was found that the QS-21 injection group showed shrinkage of muscle fibers and massive inflammatory cell infiltration, compared to the negative control group. Whereas the compounds disclosed in this disclosure only undergo slight inflammatory cell recruitment, indicating that they would be more safe than QS-21.

2. Immune potentiation of recombinant hepatitis B virus surface antigen (HBsAg)

The following immunogenic compositions were prepared according to Table 12, and the compounds were dissolved with appropriate solvents, e.g., water, DMSO, 0.9% NaCl solution, etc. And uniformly mixing the dissolved compound with the antigen.

Table 12

The results of the antibody titres are shown in figure 6, and the analysis shows that the above compounds can induce strong antibody titres except saikosaponin E when mixed with HBsAg, which is far better than that of an antigen group alone, wherein the antibody titres induced by 5 mug aescine Ie are equivalent to those induced by 25 mug aluminum adjuvant, and the two are not significantly different.

3. Immune potentiation against the H1N1 influenza virus lytic subunit antigen (H1N 1)

The following immunogenic compositions were prepared according to Table 13, and the compounds were dissolved with a suitable solvent, for example, water, DMSO, 0.9% NaCl solution, etc. And uniformly mixing the dissolved compound with the antigen.

TABLE 13

Babl/c mice with the age of 6-8 weeks are randomly divided into 5 groups of 5 mice each. Two immunizations (5. Mu.g of compound, 0.3. Mu.g of antigen, 25. Mu.g of aluminum adjuvant) were performed at weeks 0 and 3, blood was collected at week 5, and then blood-inhibitory titers were determined using fresh chicken blood.

The results are shown in FIG. 7, where 5 μg of polygalasaponin D, saikosaponin B2 induced neutralizing antibody titres better than 25 μg of aluminum adjuvant induced levels when used as influenza lyses Miao Zuoji, were all much higher than the antigen group alone.

4. Immune potentiation of novel coronavirus Omicron strain recombinant S-trimer

The following immunogenic compositions were prepared according to Table 14, and the compounds were dissolved with appropriate solvents, e.g., water, DMSO, 0.9% NaCl solution, etc. And uniformly mixing the dissolved compound with the antigen.

TABLE 14

The same batch of Babl/c mice with the age of 6-8 weeks is taken and randomly divided into 6 groups of 5 mice. Two immunizations (5. Mu.g of compound, 0.3. Mu.g of antigen, 25. Mu.g of aluminum adjuvant) were performed at weeks 0 and 2, blood was collected at week 3, and then IgG antibody titer was determined.

The analysis result is shown in fig. 8, and the result shows that several small molecule compounds disclosed in the patent have good promotion effect on the exertion of the immune effect of the antigen, and compared with the single antigen group, the antibody titer can be obviously improved (the average value is improved by more than 100 times).

The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.

Claims

1. The use of a compound or a pharmaceutically acceptable derivative thereof for the preparation of a non-specific immune response enhancer, characterized in that the compound is represented by formula I,

wherein,

R ₁ independently selected from H, a substitutable aryl or heteroaryl group;

R ₂ independently selected from H, a substitutable aryl or heteroaryl group;

R ₃ independently selected from H, a substitutable aryl or heteroaryl group;

R ₄ is H or OH;

R ₅ h or OH.

2. The use according to claim 1, wherein the compound is a saponin compound.

3. The use according to claim 1, wherein R is ₁ Is independently selected from H,

The R is ₂ Is independently selected from H,

The R is ₃ Independently selected from H, OH or COH.

4. Use according to claim 1, characterized in that the compound is selected from the following compounds:

5. use according to claim 1, characterized in that the compound is selected from the following compounds:

6. the use according to claim 1, wherein the pharmaceutically acceptable derivative is in the form of a pharmaceutically acceptable acid or basic salt thereof;

preferably, the acid salt is a hydrochloride, sulfate, phosphate, citrate, hydrobromide, acetate, benzoate, benzenesulfonate, tartrate, carbonate, citrate, gluconate, lactate, malate, methanesulfonate, stearate, valerate or nitrate; the basic salt is sodium salt, calcium salt, potassium salt, zinc salt or meglumine salt.

7. The use according to claim 1, wherein the compound further comprises a form of an isomer, hydrate, solvate, metabolite or prodrug thereof.

8. Use of a compound of formula 1 as defined in claim 1 or a pharmaceutically acceptable derivative thereof for the preparation of an immunogenic or pharmaceutical composition.

9. The use according to claim 8, wherein the immunogenic or pharmaceutical composition is a composition comprising an inactivated vaccine, an attenuated live vaccine, a protein vaccine, a bacterial polysaccharide and polysaccharide protein conjugate vaccine, a genetically engineered vaccine or a genetically reassortant vaccine.

10. A vaccine composition, characterized in that the vaccine composition comprises an immunogenic substance and a compound represented by formula 1 or a pharmaceutically acceptable derivative thereof as claimed in claim 1.

11. A compound adjuvant comprising a compound represented by formula 1 or a pharmaceutically acceptable derivative thereof as claimed in claim 1, and other non-specific immune response enhancers;

preferably, the other nonspecific immune response enhancer is one or more selected from aluminum adjuvant, calcium adjuvant, liposome, MPL, MF-59, SAF, freund's adjuvant, saponin adjuvant, manganese salt adjuvant, AS03, AS04, tween 80, bacterial lipopolysaccharide adjuvant, RIBI adjuvant system, cpG-ODN adjuvant, and imidazopyridine compound.