American ginseng root parts, methods of making and uses thereof

Abstract

The present application relates to fractions from the root portion of the plant Panax quinquefolius, and in particular to an intermediate polysaccharide fraction, a neutral polysaccharide fraction, an acid polysaccharide-enriched fraction, and an acid polysaccharide fraction, as well as methods of making them, compositions comprising them, and their use in therapy. For example, the fractions of the present application can be used to treat diseases, disorders, or conditions that are treatable by activating pattern recognition receptors, such as Toll-like receptors (TLRs), or by activating innate and adaptive immune responses, such as viral infections.

Description

[0001] Related applications

[0002] This application claims priority to co-pending U.S. provisional patent application No. 63 / 125,607, filed on December 15, 2020, the contents of which are incorporated herein by reference in their entirety. Technical Field

[0003] This application relates to fractions derived from the root portion of the American ginseng (Panax quinquefolius) plant. Specifically, the invention relates to neutral polysaccharide fractions, acid-rich polysaccharide fractions, and acidic polysaccharide fractions, as well as methods for their preparation, compositions comprising them, and their uses in treatment. Specifically, the invention relates to fractions derived from the root portion of the American ginseng (Panax quinquefolius) plant and their use in treating diseases, disorders, or conditions (e.g., viral infections) treatable by activating innate and adaptive immune responses. Background Technology

[0004] The mammalian immune system has evolved a multi-layered, layered, and interactive defense system to prevent infection. These are broadly categorized into innate immunity and adaptive immunity. Innate immunity is the first line of defense against microbial pathogens, functioning almost immediately by activating phagocytes and antigen-presenting cells (APCs), such as dendritic cells (DCs) and macrophages, to limit the early proliferation and spread of infectious agents. It also triggers an inflammatory response and defense by releasing a variety of cytokines, chemokines, and antimicrobial factors (such as interferon (IFN)). While evolutionarily ancient, innate immunity has been largely neglected by immunologists for many years due to its relative nonspecificity. However, in most cases, our innate immune system protects us from infection. On the other hand, if an infectious organism penetrates the innate immune defenses, our innate defenses promote and guide the generation of adaptive immune responses that target highly specific determinants uniquely expressed by the invading pathogen. These responses depend on rearrangements of specific antigen receptor genes in B cells and T cells, leading to the production of high-affinity antigen-specific antibodies (humoral immunity) and T cell or cell-mediated immunity. Generally, antibodies help clear, destroy, or neutralize extracellular pathogens and their toxins, while T cell-mediated immune responses help eliminate or control intracellular pathogens. Compared to innate immune responses, adaptive immune responses are characterized by specific immune memory.

[0005] Until recently, key unresolved questions concerned how the host's innate immune system detects infection and how it distinguishes itself from pathogens or infectious non-self pathogens. The discovery and characterization of the Toll-like receptor (TLR) family provided profound insights into innate immune recognition and established the crucial role of the innate immune system in host defense against infection (Akira et al., 2006; Hargreaves and Medzhitov, 2005; Kawai and Akira, 2006; Philpott and Girardin, 2004; Seth et al., 2006). The innate immune system uses multiple germline-encoded pattern recognition receptor (PRR) families to detect infection and trigger various antimicrobial defense mechanisms (Janeway and Medzhitov, 1998). These PRRs are highly conserved evolutionarily, ranging from plants and fruit flies to mammals. Innate immune recognition strategies are based on the detection of highly conserved and essential structures present in many microorganisms but absent in host cells (Janeway, 1992; Janeway and Medzhitov, 1999). Because innate immune recognition targets conserved molecular patterns, these are called pathogen-associated molecular patterns (PAMPs). PAMPs possess three important characteristics that make them ideal targets for innate immune sensing. First, PAMPs are produced solely by microorganisms, not by host cells. This is fundamental to distinguishing self-infectious pathogens from non-self-infectious pathogens. Second, PAMPs are conserved across a given class of microorganisms. This allows a limited number of PRRs to detect the presence of a large class of invading pathogens. For example, patterns in lipopolysaccharide (LPS) allow a single PRR to detect the presence of any Gram-negative bacteria. Third, PAMPs are essential for microbial survival; any mutation or loss of a PAMP is either lethal to the organism or significantly reduces its fitness. These new insights into innate immune recognition are revolutionizing our understanding of immune defense, pathogenesis, and the treatment and prevention of infectious diseases.

[0006] Toll-like receptors (TLRs) are prorheological receptors (PRRs) that recognize PAMPs. TLR signaling is highly complex and has been reviewed elsewhere (Akira and Takeda, 2004; O'Neill, 2006). In short, all TLRs except TLR3 are signaled via the adaptor molecule myeloid differentiation factor 88 (MyD88), a cytoplasmic protein containing both a Toll-IL-1 receptor (TIR) ​​domain and a death domain. Ultimately, NF-κB and MAPK are activated downstream of TRAF6, leading to the production of pro-inflammatory cytokines and chemokines such as TNF-α, IL-6, IL-1β, and IL-12. In addition to MyD88, TLR3 and TLR4 are also signaled via TRIF, another TIR-containing adaptor required for the production of type I interferon and type I interferon-dependent genes. TLRs are expressed on a variety of immune and non-immune cells. Mouse macrophages express TLRs 1–9, reflecting their importance in the initiation of pro-inflammatory responses. Plasma cell-like dendritic cells (pDCs) produce large amounts of type I interferon during viral infection and express TLR7 and 9. In mice, all conventional dendritic cells (DCs) express TLR1, 2, 4, 6, 8, and 9, while TLR3 is limited to the CD8+ and CD4-CD8-DC subsets (Iwasaki and Medzhitov, 2004). In humans, TLR9 expression is limited to pDCs and B cells (Bauer et al., 2001; Krug et al., 2001). TLR3 is not expressed in mouse or human pDCs. In addition to focusing on expression on immune cells, there is also interest in understanding TLR expression on mucosal epithelial cells (ECs), which form the first line of defense against most infections.

[0007] TLR induces a range of responses, depending on the cell type in which they are activated (Ashkar and Rosenthal, 2002; Iwasaki and Medzhitov, 2004). For example, treatment of dendritic cells (DCs) with CpG DNA that functions via TLR9 activates DC maturation, including upregulation of class II MHC and co-stimulatory molecules, as well as the production of pro-inflammatory cytokines, chemokines, and enhanced antigen presentation. Similarly, treatment of B cells with CpG induces their activation and proliferation, secretion of antibodies as well as IL-6 and IL-10, and resistance to apoptosis. Activation of immune cells via CpG DNA induces a Th1-dominated response.

[0008] The mechanisms by which PRRs mediate host defense against pathogens are a focus of in-depth research. Due to their ability to enhance innate immune responses, novel strategies using PRR ligands, natural or synthetic agonists or antagonists—the so-called “innate immunology”—can be used alone or in combination with other antiviral agents to provide protection or treatment against intracellular bacterial, parasitic, and viral infections. Furthermore, activating the innate immune system via PRRs (e.g., TLRs) using their respective ligands or agonists represents a strategy to enhance immune responses against specific pathogens, making PRR-mediated signaling an excellent vaccine adjuvant.

[0009] Korean red ginseng has been shown to have antiviral effects through immunomodulation (Nguyen and Nguyen, 2019). Summary of the Invention

[0010] The applicant has developed a method for preparing polysaccharide extracts and fractions from the roots of the American ginseng (Panax quinquefolius) plant. Specifically, the applicant has produced intermediate polysaccharide fractions, from which neutral polysaccharide fractions, acid-rich polysaccharide fractions, and acidic polysaccharide fractions have been produced.

[0011] Therefore, this application includes a method for preparing intermediate polysaccharide fractions from the root of the American ginseng (Panax quinquefolius) plant, the method comprising:

[0012] At temperatures ranging from approximately 10°C to approximately 90°C, using a solution containing approximately 50% (v / v) to approximately 95% (v / v) C 1-3 A solvent mixture of alkyl OH and water is used to extract the root portion from the root portion of Panax quinquefolius to produce a solvent mixture fraction and a first residue, wherein the ratio of the solvent mixture to the root portion is about 20 to about 1 (v / w) to about 1 to about 1 (v / w).

[0013] The first residue is fractionated from the solvent mixture;

[0014] The first residue is extracted with water at a temperature of about 10°C to about 100°C to produce an aqueous fraction comprising the intermediate polysaccharide fraction and a second residue, wherein the ratio of water to the root fraction is about 20 (v / w) to about 1 (v / w) to about 1 (v / w) to about 1 (v / w); and

[0015] The aqueous fraction containing the intermediate polysaccharide fraction is separated from the second residue; and

[0016] Optionally, the aqueous fraction is dried or concentrated to produce the intermediate polysaccharide fraction.

[0017] This application also includes a method for preparing neutral polysaccharide fractions, the method comprising:

[0018] Provide intermediate polysaccharide fractions produced by the above method;

[0019] The intermediate polysaccharide fraction is dissolved in water to produce a solution of the intermediate polysaccharide fraction, wherein the solution of the intermediate polysaccharide fraction is from about 1% (w / w) to about 10% (w / w);

[0020] C 1-3 Alkyl OH groups are added to the solution of the intermediate polysaccharide fraction to produce a neutral polysaccharide fraction and a first supernatant, wherein the C 1-3 The ratio of the alkyl OH to the intermediate polysaccharide fraction solution is about 1 to about 1 (v / v) to about 0.5 to about 1 (v / v);

[0021] The neutral polysaccharide fraction is separated from the first supernatant to produce the neutral polysaccharide fraction; and

[0022] Alternatively, in addition to using the neutral polysaccharide fraction, the neutral polysaccharide fraction can be further purified by repeating the preceding three steps.

[0023] This application includes a method for preparing acid-rich polysaccharide fractions, the method comprising:

[0024] Provide intermediate polysaccharide fractions produced by the above method;

[0025] The intermediate polysaccharide fraction is dissolved in water to produce a solution of the intermediate polysaccharide fraction, wherein the solution of the intermediate polysaccharide fraction (PBG-002) is from about 1% (w / w) to about 10% (w / w).

[0026] C 1-3 Alkyl OH groups are added to the solution of the intermediate polysaccharide fraction to produce a neutral polysaccharide fraction and a first supernatant, wherein the C 1-3 The ratio of the alkyl OH to the intermediate polysaccharide fraction solution is about 1 to about 1 (v / v) to about 0.5 to about 1 (v / v);

[0027] The first supernatant was separated from the neutral polysaccharide;

[0028] C 1-3 Alkyl OH is added to the first supernatant to produce a second supernatant rich in acidic polysaccharide fractions, wherein C 1-3 The ratio of alkyl OH to the first supernatant is about 0.5 to about 1 (v / v) to about 8 to about 1 (v / v);

[0029] The acid-rich polysaccharide fraction is separated from the second supernatant to produce the acid-rich polysaccharide fraction.

[0030] This application also includes a method for preparing acidic polysaccharide fractions, the method comprising:

[0031] Provides acid-rich polysaccharide fractions produced by the above method;

[0032] The acid-rich polysaccharide fraction is dissolved in water to produce a solution rich in acid-rich polysaccharide fraction, wherein the solution rich in acid-rich polysaccharide fraction is from about 1% (w / w) to about 10% (w / w);

[0033] C 1-3 Alkyl OH groups are added to the solution rich in the acidic polysaccharide fraction to produce a precipitate and a third supernatant containing the acidic polysaccharide fraction, wherein C 1-3 The ratio of alkyl OH to the solution rich in acidic polysaccharide fraction is about 1 to about 1 (v / v) to about 0.5 to about 1 (v / v);

[0034] The third supernatant containing the acidic polysaccharide fraction is separated from the precipitate; and

[0035] Optionally, the third supernatant may be dried or concentrated to produce the acidic polysaccharide fraction.

[0036] Alternatively, the acid-rich polysaccharide fraction can be purified using column chromatography to produce the acidic polysaccharide fraction. Therefore, this application also includes a method for preparing the acidic polysaccharide fraction, the method comprising:

[0037] Provides acid-rich polysaccharide fractions produced by the above method;

[0038] The acid-rich polysaccharide fraction is fractionated from a chromatographic column containing diethylaminoethyl (DEAE) anion exchange resin to produce an acidic polysaccharide elution fraction, wherein the fractionation includes:

[0039] (i) Dissolve the acid-rich polysaccharide fraction in water to produce a solution rich in acid-rich polysaccharide fraction;

[0040] (ii) Loading the solution rich in acidic polysaccharide fractions onto a chromatographic column containing the anion exchange resin; and

[0041] (iii) Eluting the acid-rich polysaccharide-rich elution fraction from the anion exchange resin using ammonium acetate buffer to produce the acid-rich polysaccharide elution fraction, wherein the elution comprises a first step in which the buffer is about 10 mM to about 100 mM ammonium acetate, and a second step in which the buffer is about 0.5 M to about 1.5 M ammonium acetate; and

[0042] Optionally, the acidic polysaccharide elution fraction can be dried or concentrated to produce the acidic polysaccharide fraction.

[0043] This application also includes intermediate polysaccharide fractions produced by the above method.

[0044] In one embodiment, the present application includes an intermediate polysaccharide fraction containing about 0.5 mol% to about 4 mol% rhamnose (Rha), about 13 mol% to about 17 mol% galacturonic acid (GalA), about 70 mol% to about 74 mol% glucose (Glc), about 3 mol% to about 7 mol% galactose (Gal) and about 3 mol% to about 7 mol% arabinose (Ara).

[0045] In one embodiment, this application also includes neutral polysaccharide fractions produced by the above method.

[0046] This application also includes a neutral polysaccharide fraction containing about 0.1 mol% to about 3 mol% rhamnose (Rha), about 3 mol% to about 7 mol% galacturonic acid (GalA), about 87 mol% to about 92 mol% glucose (Glc), about 1 mol% to about 4 mol% galactose (Gal) and about 1 mol% to about 5 mol% arabinose (Ara).

[0047] In one embodiment, the present application also includes acid-rich polysaccharide fractions produced by the above method.

[0048] In one embodiment, the present application further includes acidic polysaccharide fractions produced by the above method.

[0049] This application includes an acidic polysaccharide fraction containing about 2 mol% to about 6 mol% rhamnose (Rha), about 43 mol% to about 47 mol% galacturonic acid (GalA), about 10 mol% to about 14 mol% glucose (Glc), about 18 mol% to about 22 mol% galactose (Gal) and about 16 mol% to about 20 mol% arabinose (Ara).

[0050] This application also includes a composition comprising one or more fractions and a carrier of this application.

[0051] This application includes a method for activating the function of Toll-like receptors (TLRs) in biological samples or patient cells, comprising administering an effective amount of one or more fractions of this application to the cells.

[0052] This application further includes a method for stimulating the production of one or more chemokines in biological samples or patient cells, comprising administering an effective amount of one or more fractions of this application to the cells.

[0053] This application also includes a method for treating a disease, disorder, or condition by activating TLR function, the method comprising administering a therapeutically effective amount of one or more fractions of this application to a subject in need of such treatment.

[0054] This application also includes a method for activating innate and / or adaptive innate responses in biological samples or patient cells, comprising administering an effective amount of one or more fractions of this application to the cells.

[0055] This application also includes a method for treating a viral infection, comprising administering a therapeutically effective amount of one or more fractions of this application to a subject in need of it.

[0056] Other features and advantages of this application will become apparent from the following detailed description. However, it should be understood that while the detailed description and specific examples indicate embodiments of this application, they are given by way of illustration only, and the scope of the claims should not be limited by these embodiments, but should be given the broadest interpretation consistent with the entire specification.

[0057] Brief description of the attached figures

[0058] The embodiments of this application will now be described in more detail with reference to the accompanying drawings, wherein:

[0059] Figure 1 This is a flowchart showing the preparation process of the intermediate polysaccharide fraction (PBG-002) and the purification process of the resulting neutral polysaccharide fraction (PBG-005), acid-rich polysaccharide fraction (PBG-003), and acidic polysaccharide fraction (PBG-007).

[0060] Figure 2 The cells expressing TLR2 showed that they stimulated the production of IL-8.

[0061] Figure 3 The cells expressing TLR4 stimulate the production of IL-8.

[0062] Figure 4A The effect of the fractions applied in this application on H1N1 virus titers in macrophage cultures was shown. *p<0.05, **p<0.001. Figure 4B The effect of the fractions applied in this application on H1N1 virus titers in macrophage cultures was shown. *p<0.05, **p<0.001.

[0063] Figure 5 The effect of the fractions applied in this application on HSV-1 viral titers in mouse dermal fibroblast cultures was shown. *p<0.05, **p<0.001.

[0064] Figure 6The effect of the fractions applied in this application on HSV-1 viral titers in human skin fibroblast cell culture was shown. *p<0.05, **p<0.001.

[0065] Figure 7 The effect of the fractions applied in this application on HSV-2 viral titers in mouse dermal fibroblast cultures was shown. *p<0.05, **p<0.001.

[0066] Figure 8 The effect of the fractions applied in this application on HSV-2 viral titers in human skin fibroblast cell culture was shown. *p<0.05, **p<0.001.

[0067] Figure 9A and Figure 9B The effect of the fractions applied in this application on HSV-2 viral titers in human skin fibroblast cell culture was shown. *p<0.05, **p<0.001.

[0068] Figure 10 The effect of the fractions used in this application on the viral titer in guinea pig vaginal washes following an HSV-2 IVAG attack was demonstrated.

[0069] Figure 11 This demonstrates the impact of the application's rating on vaginal lesion scores during initial HSV-2 infection.

[0070] Figure 12A This study demonstrates the impact of the application's scoring system on the total cumulative recurrent vaginal lesion score following primary HSV-2 infection. Figure 12B This study demonstrates the impact of the application's grading system on the cumulative number of recurrent vaginal lesions in individuals following primary HSV-2 infection.

[0071] Figure 13 The effect of the application rating on the survival percentage of guinea pigs after an HSV-3 IVAG attack is shown.

[0072] Figure 14 The effect of the application rating on the daily body weight of guinea pigs after HSV-2 IVAG attack was shown. Specific Implementation

[0073] 1. Definition

[0074] Unless otherwise stated, those skilled in the art will understand that the definitions and embodiments described in this section and other parts are intended to be applicable to all embodiments and aspects of this application to which they are suited.

[0075] As used herein, the terms “fraction of the application” or “fraction of this application” refer to intermediate polysaccharide fraction (PBG-002), neutral polysaccharide fraction (PBG-005), acid-rich polysaccharide fraction and / or acid polysaccharide fraction or their enantiomers, salts and / or solvates.

[0076] As used herein, the terms “composition of the application” or “composition of this application” refer to compositions comprising one or more fractions of this application.

[0077] As used herein, the term "and / or" means that the listed items are present or used individually or in combination. In practice, the term implies the use or presence of "at least one" or "one or more" of the listed items. The term "and / or" with respect to their salts and / or solvates means that the application of each fraction of this application is present as a salt and hydrate individually, or in combination thereof, such as a salt of a solvate of a compound of this application.

[0078] As used herein, the singular forms “a,” “an,” and “the” include plural references unless the content explicitly indicates otherwise. For example, embodiments including “negative response” should be understood to present one negative response, or two or more additional negative responses, in some respects.

[0079] In embodiments that include an “additional” or “second” component or effect (e.g., an additional or second adverse effect), the second effect as used herein is different from other effects or the first effect. A “third” adverse effect is different from other, first and second effects, and similarly different from the further enumerated or “additional” adverse effects.

[0080] As used herein, the term “suitable” means that the selection of a particular compound or condition will depend on the specific synthetic operation to be performed, the properties of the molecule to be transformed, and / or the specific use of the compound, but that the selection is entirely within the skill of a person trained in the art.

[0081] As used herein, the terms “about,” “substantially,” and “approximately” refer to a reasonable amount of deviation of the modified term such that the final result is not significantly altered. These degree terms should be interpreted as including at least ±5% deviation of the modified term, assuming that such deviation does not negate the meaning of the word it modifies, unless the context otherwise implies to those skilled in the art.

[0082] In embodiments containing an “additional” or “second” component, the second component as used herein is chemically different from the other components or the first component. A “third” component is different from the other, first and second components, and similarly different from the further listed or “additional” components.

[0083] The term "salt" refers to an acid addition salt or a base addition salt. The term "salt" includes salts commonly used to form addition salts of free acids or free bases, as well as those salts compatible with plant treatments.

[0084] As used herein, the term "solvent" refers to a compound or salt of a compound in which molecules of a suitable solvent are incorporated into the crystal lattice. A suitable solvent is physiologically tolerable at the administered dose.

[0085] As used herein, the term "disease, disorder, or condition associated with activation of innate and / or adaptive innate responses" refers to any disease, disorder, or condition that can be treated directly or indirectly by activation of innate and / or adaptive innate responses.

[0086] As used herein, the term "subject" includes all members of the animal kingdom, including mammals, and appropriately refers to humans. Therefore, the methods and uses of this application are suitable for human treatment and veterinary applications.

[0087] As used in this article, the term "pharmaceutical composition" refers to a composition of substances intended for pharmaceutical use.

[0088] The term "pharmaceutically acceptable salt" refers to an acid addition salt or base addition salt that is suitable for or compatible with the treatment of the subject.

[0089] The term "pharmaceutically acceptable salt" includes salts that are commonly used to form addition salts of free acids or free bases.

[0090] As used herein, the term "effective dose" or "therapeutic effective dose" refers to a dose that is effective both in terms of dosage and in terms of the time period required to achieve the desired results.

[0091] As used herein, the terms “treat,” “treating,” and “treatment” are well-known in the art and refer to methods for achieving beneficial or desired outcomes, including clinical outcomes. Examples of beneficial or desired clinical outcomes include, but are not limited to: reducing the severity of a viral or microbial infection; stabilizing (i.e., not worsening) the state of a viral or microbial infection; preventing the spread of a viral or microbial infection; delaying or slowing the progression of an infection; improving or alleviating the state of a viral or microbial infection; reducing the recurrence of a viral infection; reducing, stabilizing, alleviating, or improving one or more diseases, disorders, or conditions caused by a viral or microbial infection; reducing the recurrence of one or more diseases, disorders, or conditions caused by a viral or microbial infection; and alleviating a viral or microbial infection and / or one or more symptoms or conditions caused by a viral or microbial infection, whether partial or complete, detectable or undetectable. “Treatment,” “treatment,” and “treatment” can also mean prolonged survival compared to expected survival without treatment. As used herein, “treatment,” “treatment,” and “treatment” also include preventative treatment. For example, treating a subject with an early viral or microbial infection to prevent progression, or treating a subject in remission to prevent recurrence.

[0092] "Mitigation" of infection, disease, disorder, and / or condition refers to a reduction in the severity and / or undesirable clinical manifestations and a slower or longer progression of the infection, disease, disorder, and / or condition compared to no treatment.

[0093] As used in this article, “viral infection” refers to the invasion of cells or body tissues by one or more foreign, unwanted viruses.

[0094] The term "fraction" refers to a concentrated preparation of plant material produced by extracting or precipitating plant material with a suitable solvent or solvent mixture.

[0095] The term "intermediate fraction" refers to a concentrated preparation of plant material produced by extraction or precipitation of plant material with a suitable solvent or solvent mixture, from which additional "fractions" can be generated.

[0096] As used in this article, the terms “nutritional products” or “functional foods” refer to natural bioactive chemical compounds that provide physiological benefits (such as disease prevention or treatment and / or health promotion) and can be used to supplement a diet.

[0097] As used herein, the term "PBG-002" refers to the intermediate polysaccharide fraction of this application.

[0098] As used herein, the term "PBG-005" refers to the neutral polysaccharide fraction of this application.

[0099] As used herein, the term "PBG-003" refers to the acid-rich polysaccharide fraction of this application.

[0100] As used herein, the terms “PBG-007”, “PBG-007-1”, or “PBG-007-2” refer to the acidic polysaccharide fraction of this application.

[0101] When used, for example, in relation to the treatment methods, uses, compositions and kits of this application, the subject, such as the subject who is “in need”, is a subject who is diagnosed with, suspected of having, may have been exposed to, and / or has previously received treatment for a disease, disorder or condition caused by a viral or microbial infection.

[0102] 2. Method for preparing the fractions of this application

[0103] The applicant has developed a method for preparing and obtaining polysaccharide extracts or fractions from the roots of the American ginseng (Panax quinquefolius) plant. Specifically, the applicant obtained an intermediate polysaccharide fraction (PBG-002), from which a neutral polysaccharide fraction (PBG-005), an acid-rich polysaccharide fraction (PBG-003), and an acidic polysaccharide fraction (PBG-007) were obtained. The intermediate, neutral, acid-rich, and acidic polysaccharide fractions have been shown to activate Toll-like receptors (TLRs) and stimulate the production of one or more chemokines. Therefore, the intermediate, neutral, acid-rich, and acidic polysaccharide fractions have been demonstrated to be TLR agonists and immunomodulators, such as immunostimulants, which can activate the innate and adaptive immune systems, particularly the innate immune system. Intermediate polysaccharide fractions, neutral polysaccharide fractions, acid-rich polysaccharide fractions, and acidic polysaccharide fractions have also been shown to have significant beneficial effects on viral titers, and thus have been demonstrated to provide protection against viral infection in both in vitro and in vivo models. Therefore, intermediate polysaccharide fractions, neutral polysaccharide fractions, acid-rich polysaccharide fractions, and acidic polysaccharide fractions have shown antiviral activity.

[0104] Therefore, this application includes a method for preparing an intermediate polysaccharide fraction (PBG-002) from the root of the American ginseng (Panax quinquefolius) plant, the method comprising:

[0105] At temperatures ranging from approximately 10°C to approximately 90°C, using a solution containing approximately 50% (v / v) to approximately 95% (v / v) C 1-3 A solvent mixture of alkyl OH and water was used to extract the root portion from the root portion of the Panax quinquefolius plant to produce a solvent mixture fraction and a first residue, wherein the ratio of the solvent mixture to the root portion was about 20 to about 1 (v / w) to about 1 to about 1 (v / w).

[0106] The first residue is fractionated from the solvent mixture;

[0107] The first residue is extracted with water at a temperature of about 10°C to about 100°C to produce an aqueous fraction comprising the intermediate polysaccharide fraction and a second residue, wherein the ratio of water to the root fraction is about 20 to about 1 (v / w) to about 1 to about 1 (v / w); and

[0108] The aqueous fraction containing the intermediate polysaccharide fraction is separated from the second residue; and

[0109] Optionally, the aqueous fraction is dried or concentrated to produce the intermediate polysaccharide fraction.

[0110] Those skilled in the art will understand that the root portion is any part of the plant root. In one embodiment, the root portion is a fresh root portion. In one embodiment, the root portion is a dried root portion. In one embodiment, the dried root portion is white ginseng. In one embodiment, the dried root portion is red ginseng.

[0111] In one embodiment, the root portion is pulverized before extraction to produce a pulverized root portion. Therefore, the root portion is a pulverized root portion. In one embodiment, the pulverized root portion is micronized. Therefore, the pulverized root portion is a micronized root portion. In one embodiment, the micronized root portion is micronized by milling, hammering, or grinding. In one embodiment, the root portion is micronized by grinding. Therefore, in one embodiment, the root portion is a milled root portion. In one embodiment, the root portion is a pulverized root portion.

[0112] In one embodiment, the root portion may be obtained globally from multiple commercial sources. Commercial sources include, for example, department stores, roadside shops, large retailers, supermarkets, duty-free shops, pharmacies, traditional markets, and online retail sources.

[0113] In one embodiment, the particle size range of the root portion is about 100 μm to about 1500 μm, about 100 μm to about 1000 μm, about 150 μm to about 900 μm, about 150 μm to about 800 μm, about 200 μm to about 600 μm, or about 200 μm to about 450 μm.

[0114] In one embodiment, during the step of extracting the root portion, the solvent mixture contains approximately 55% C. 1-3 Alkyl OH (v / v) to approximately 95% C 1-3 Alkyl OH (v / v), approximately 60% C 1-3 Alkyl OH (v / v) to approximately 95% C1-3 Alkyl OH (v / v), approximately 70% C 1-3 Alkyl OH (v / v) to approximately 95% C 1-3 Alkyl OH (v / v), approximately 75% C 1-3 Alkyl OH (v / v) to about 90% C 1-3 Alkyl OH (v / v), approximately 80% C 1-3 Alkyl OH (v / v) to about 90% C 1-3 Alkyl OH (v / v), approximately 80% C 1-3 Alkyl OH (v / v) to about 85% C 1-3 Alkyl OH (v / v). In one embodiment, the solvent mixture contains about 60% C 1-3 Alkyl OH (v / v) to approximately 95% C 1-3 Alkyl OH (v / v), approximately 75% C 1-3 Alkyl OH (v / v) to about 90% C 1-3 Alkyl OH (v / v) or about 80% C 1-3 Alkyl OH (v / v) to about 90% C 1-3 Alkyl OH (v / v). In one embodiment, the solvent mixture contains about 80% C 1-3 Alkyl OH (v / v) to about 90% C 1-3 Alkyl OH (v / v). In one embodiment, the solvent mixture contains about 50% C 1-3 Alkyl OH (v / v), approximately 60% C 1-3 Alkyl OH (v / v), approximately 70% C 1-3 Alkyl OH (v / v), approximately 75% C 1-3 Alkyl OH (v / v), approximately 80% C 1-3 Alkyl OH (v / v), approximately 85% C 1-3 Alkyl OH (v / v), approximately 90% C 1-3 Alkyl OH (v / v) or about 95% C 1-3 Alkyl OH (v / v). In one embodiment, the solvent mixture contains about 75% C 1-3 Alkyl OH (v / v), approximately 80% C 1-3 Alkyl OH (v / v), approximately 85% C 1-3 Alkyl OH (v / v), approximately 90% C 1-3 Alkyl OH (v / v) or about 95% C 1-3 Alkyl OH (v / v). In one embodiment, the solvent mixture contains about 85% C 1-3 Alkyl OH (v / v). In one embodiment, the solvent mixture contains more than about 75% C. 1-3 Alkyl OH (v / v), greater than about 80% C1-3 Alkyl OH (v / v), greater than about 85% C 1-3 Alkyl OH (v / v), greater than about 90% C 1-3 Alkyl OH (v / v) or greater than about 95% C 1-3 Alkyl OH (v / v). In one embodiment, the solvent mixture contains more than about 85% C. 1-3 Alkyl OH (v / v).

[0115] In one embodiment, during the step of extracting the root portion, the C 1-3 The alkyl OH group is selected from methanol, ethanol, propanol, and isopropanol. In one embodiment, C 1-3 The alkyl OH is selected from methanol and ethanol. In one embodiment, C 1-3 The alkyl OH group represents ethanol.

[0116] In one embodiment, during the step of extracting the root portion, the ratio of the solvent mixture to the root portion is about 18:about 1 (v / w), about 15:about 1 (v / w), about 10:about 1 (v / w), about 5:about 1 (v / w), or about 1:about 1 (v / w). In one embodiment, during the step of extracting the root portion, the ratio of the solvent mixture to the root portion is about 15:about 1 (v / w) to about 5:about 1 (v / w). In one embodiment, during the step of extracting the root portion, the ratio of the solvent mixture to the root portion is about 10:about 1 (v / w). In one embodiment, the (v / w) is L / Kg.

[0117] In one embodiment, the step of extracting the root portion with a solvent mixture is carried out at the following temperatures: about 20°C to about 90°C, about 30°C to about 90°C, about 40°C to about 90°C, about 50°C to about 90°C, about 50°C to about 60°C, about 50°C to about 70°C, about 50°C to about 80°C, about 60°C to about 70°C, about 60°C to about 80°C, about 70°C to about 80°C, about 50°C to about 90°C, about 60°C to about 90°C, about 70°C to about 90°C, and about 80°C to about 90°C. In one embodiment, the step of extracting the root portion from the root portion of the Panax quinquefolius plant with a solvent mixture is carried out at the following temperatures: about 20°C to about 90°C, about 30°C to about 90°C, about 40°C to about 90°C, about 50°C to about 90°C, about 60°C to about 90°C, about 70°C to about 90°C, or about 80°C to about 90°C. In one embodiment, the step of extracting the root portion with a solvent mixture is performed at the boiling point of the solvent mixture.

[0118] Those skilled in the art will understand that the step of extracting the root portion with the solvent mixture should last for a certain period of time to allow sufficient extraction of unwanted substances from the first residue. In one embodiment, the step of extracting the root portion with the solvent mixture is performed for more than about 1 hour, more than about 2 hours, more than about 3 hours, more than about 4 hours, or more than about 5 hours. In one embodiment, the step of extracting the root portion with the solvent mixture is performed for more than about 1 hour, more than about 2 hours, or more than about 3 hours. In one embodiment, the step of extracting the root portion with the solvent mixture is performed for about 1 hour to about 5 hours, about 2 hours to about 3 hours, about 2 hours to about 4 hours, about 1 hour to about 4 hours, or about 1 hour to about 3 hours. In one embodiment, the step of extracting the root portion with the solvent mixture is performed for about 1 hour to about 3 hours. In one embodiment, the step of extracting the root portion with the solvent mixture is performed for about 3 hours.

[0119] In one embodiment, during the step of extracting the first residue, the water-to-root ratio is approximately 18:approximately 1 (v / w), approximately 15:approximately 1 (v / w), approximately 12:approximately 1 (v / w), approximately 10:approximately 1 (v / w), approximately 5:approximately 1 (v / w), or approximately 1:approximately 1 (v / w). In one embodiment, the water-to-root ratio is approximately 15:approximately 1 (v / w) to approximately 5:approximately 1 (v / w). In one embodiment, the ratio is approximately 12:approximately 1 (v / w).

[0120] In one embodiment, the step of extracting the first residue with water is performed at the following temperatures: about 20°C to about 100°C, about 30°C to about 100°C, about 40°C to about 100°C, about 50°C to about 100°C, about 60°C to about 100°C, about 20°C to about 90°C, about 30°C to about 90°C, about 40°C to about 90°C, about 50°C to about 90°C, about 50°C to about 60°C, about 50°C to about 70°C, about 50°C to about 80°C, about 60°C to about 70°C, about 60°C to about 80°C, about 70°C to about 80°C, about 50°C to about 90°C, about 60°C to about 90°C, about 70°C to about 90°C, about 80°C to about 100°C, about 80°C to about 100°C, or about 90°C to about 100°C. In one embodiment, the step of extracting the first residue with water is performed at about 80°C to about 100°C. In one embodiment, the step of extracting the first residue with water is performed at about 100°C.

[0121] Those skilled in the art will understand that the step of extracting the first residue with water should continue for a certain duration to allow for sufficient extraction of the intermediate polysaccharide fraction. In one embodiment, the step of extracting the first residue with water is performed for more than about 1 hour, more than about 2 hours, more than about 3 hours, more than about 4 hours, or more than about 5 hours. In one embodiment, the step of extracting the first residue with water is performed for more than about 1 hour, more than about 2 hours, or more than about 3 hours. In one embodiment, the step of extracting the first residue with water is performed for about 1 hour to about 5 hours, about 2 hours to about 3 hours, about 2 hours to about 4 hours, about 1 hour to about 4 hours, or about 1 hour to about 3 hours. In one embodiment, the step of extracting the first residue with water is performed for about 1 hour to about 3 hours. In one embodiment, the step of extracting the first residue with water is performed for more than about 1 hour. In one embodiment, the step of extracting the first residue with water is performed for about 3 hours.

[0122] Those skilled in the art will understand that any separation method known in the art can be used to separate the first residue from the solvent mixture fraction and / or to separate the aqueous fraction from the second residue. In one embodiment, the aqueous fraction is separated by filtration, decantation, or centrifugation. In one embodiment, separation is performed by centrifugation.

[0123] In one embodiment, centrifugation is performed at approximately 800 rpm to approximately 1500 rpm, approximately 1000 rpm to approximately 1500 rpm, or approximately 1100 rpm to approximately 1300 rpm. In one embodiment, centrifugation is performed at approximately 1200 rpm. In one embodiment, centrifugation is performed for approximately 5 minutes to approximately 20 minutes, approximately 5 minutes to approximately 15 minutes, or approximately 8 minutes to approximately 12 minutes. In one embodiment, centrifugation is performed for approximately 10 minutes.

[0124] In one embodiment, the drying or concentration step is performed by any solvent removal method known in the art. In one embodiment, the drying or concentration step is performed by evaporation. In one embodiment, the evaporation is performed by distillation, open-air evaporation, vacuum evaporation, or rotary evaporation. In one embodiment, the evaporation is performed by rotary evaporation.

[0125] In one embodiment, the drying step includes concentrating the aqueous fraction containing the intermediate polysaccharide, followed by freeze-drying or spray drying to produce the intermediate polysaccharide. In another embodiment, the drying includes concentrating the aqueous fraction containing the intermediate polysaccharide, followed by freeze-drying to produce the intermediate polysaccharide.

[0126] Those skilled in the art will understand that in the step of optionally drying or concentrating the aqueous fraction to produce the intermediate polysaccharide fraction, "optionally" means that after the step of separating the aqueous fraction containing the intermediate polysaccharide fraction from the second residue, the aqueous fraction containing the intermediate polysaccharide fraction can be used in the produced form, or the aqueous fraction can be further dried or concentrated as described above.

[0127] In one embodiment, the intermediate polysaccharide fraction comprises a neutral polysaccharide fraction and an acid-rich polysaccharide fraction, from which an acid-rich polysaccharide fraction is further obtained. In one embodiment, the polysaccharide extract further comprises saponins and / or oligosaccharides.

[0128] In one embodiment, the intermediate polysaccharide grade produced after drying is separated into solids, oils, waxes, or gums. In one embodiment, the intermediate polysaccharide grade is separated into solids. In one embodiment, the intermediate polysaccharide grade is separated into solids, and the solid is a powder.

[0129] In one embodiment, this application further includes a method for preparing an intermediate polysaccharide fraction (PGB-002) from the root of the American ginseng (Panax quinquefolius) plant, the method comprising:

[0130] At a temperature of about 10°C to about 90°C, the root portion of Panax quinquefolius is extracted from the root portion of the plant with a solvent mixture comprising about 50% (v / v) to about 95% (v / v) ethanol and water to produce a solvent mixture fraction and a first residue, wherein the ratio of the solvent mixture to the root portion is about 20 to about 1 (v / w) to about 1 to about 1 (v / w).

[0131] The first residue is fractionated from the solvent mixture;

[0132] The first residue is extracted with water at a temperature of about 10°C to about 100°C to produce an aqueous fraction containing the intermediate polysaccharide fraction and a second residue, wherein the ratio of water to the root fraction is about 20 to about 1 (v / w) to about 1 to about 1 (v / w).

[0133] The aqueous fraction containing the intermediate polysaccharide fraction is separated from the second residue; and

[0134] Optionally, the aqueous fraction is dried or concentrated to produce the intermediate polysaccharide fraction.

[0135] The applicant also obtained a neutral polysaccharide fraction (PBG-005) and an acid-rich polysaccharide fraction (PBG-003) from the intermediate polysaccharide.

[0136] Therefore, this application also includes a method for preparing a neutral polysaccharide fraction (PBG-005), the method comprising:

[0137] Provide intermediate polysaccharide fractions produced by the above method;

[0138] The intermediate polysaccharide fraction is dissolved in water to produce a solution of the intermediate polysaccharide fraction, wherein the solution of the intermediate polysaccharide fraction is from about 1% (w / w) to about 10% (w / w);

[0139] C 1-3 Alkyl OH groups are added to the solution of the intermediate polysaccharide fraction to produce a neutral polysaccharide fraction and a first supernatant, wherein the C 1-3 The ratio of the alkyl OH to the intermediate polysaccharide fraction solution is about 1 to about 1 (v / v) to about 0.5 to about 1 (v / v);

[0140] Separate the neutral polysaccharide fraction from the first supernatant to produce the neutral polysaccharide fraction; and

[0141] Alternatively, in addition to using the neutral polysaccharide fraction, the neutral polysaccharide fraction can be further purified by repeating the preceding three steps.

[0142] In one embodiment, the intermediate polysaccharide fraction solution is about 2% (w / w) to about 10% (w / w), about 3% (w / w) to about 10% (w / w), about 4% (w / w) to about 10% (w / w), about 5% (w / w) to about 10% (w / w), about 4% (w / w) to about 9% (w / w), about 4% (w / w) to about 8% (w / w), about 4% (w / w) to about 7% (w / w), about 4% (w / w) to about 6% (w / w); about 2% (w / w), about 3% (w / w), about 4% (w / w), about 5% (w / w), or about 6% (w / w). In one embodiment, the intermediate polysaccharide fraction solution is about 5% (w / w).

[0143] In one embodiment, C is added to the solution of the intermediate polysaccharide fraction. 1-3 The C in the alkyl OH step 1-3 The alkyl OH group is selected from methanol, ethanol, propanol, and isopropanol. In one embodiment, the C 1-3 The alkyl OH group is selected from methanol and ethanol. In one embodiment, the C 1-3 The alkyl OH group represents ethanol.

[0144] In one embodiment, C 1-3 The ratio of the alkyl OH to the intermediate polysaccharide fraction solution is about 1 to about 1 (v / v) to about 0.5 to about 1 (v / v). In one embodiment, C 1-3The ratio of the alkyl OH to the intermediate polysaccharide fraction solution is about 1 to about 1 (v / v) to about 0.5 to about 1 (v / v) or about 0.7 to about 1 (v / v) to about 0.5 to about 1 (v / v). In one embodiment, C 1-3 The ratio of the alkyl OH to the intermediate polysaccharide fraction solution is about 0.7 to about 1 (v / v) to about 0.5 to about 1 (v / v). In one embodiment, C 1-3 The ratio of the alkyl OH to the intermediate polysaccharide fraction solution is about 0.5 to about 1 (v / v).

[0145] In one embodiment, C 1-3 The step of adding alkyl OH to the solution of the intermediate polysaccharide fraction to produce a neutral polysaccharide fraction and a first supernatant is performed for more than about 1 hour, more than about 2 hours, more than about 3 hours, more than about 4 hours, more than about 5 hours, or more than about 6 hours. In one embodiment, C 1-3 The step of adding alkyl OH to the solution of the intermediate polysaccharide fraction to produce a neutral polysaccharide fraction and a first supernatant is performed for more than about 1 hour, more than about 2 hours, or more than about 3 hours. In one embodiment, C... 1-3 The step of adding alkyl OH to the solution of the intermediate polysaccharide fraction to produce a neutral polysaccharide fraction and a first supernatant is carried out for about 1 hour to about 5 hours, about 2 hours to about 3 hours, about 2 hours to about 4 hours, about 1 hour to about 4 hours, or about 1 hour to about 3 hours. In one embodiment, C 1-3 The step of adding alkyl OH to the solution of the intermediate polysaccharide fraction to produce a neutral polysaccharide fraction and a first supernatant is carried out for about 1 hour to about 3 hours. In one embodiment, C 1-3 The step of adding alkyl OH to the solution of the intermediate polysaccharide fraction to produce a neutral polysaccharide fraction and a first supernatant is performed in the C... 1-3 The alkyl OH group was added to the solution of the intermediate polysaccharide fraction and carried out for approximately 3 hours. In one embodiment, C... 1-3 The step of adding alkyl OH to the solution of the intermediate polysaccharide fraction to produce a neutral polysaccharide fraction and a first supernatant is carried out for more than about 1 hour.

[0146] In one embodiment, in addition to using the neutral polysaccharide fraction to produce a purified neutral polysaccharide fraction, the neutral polysaccharide fraction may optionally be further purified by repeating the first three steps of the above method. In one embodiment, the neutral polysaccharide fraction may optionally be further purified by repeating the step of dissolving the intermediate polysaccharide fraction in water to the step of separating the neutral polysaccharide fraction from the first supernatant to produce the neutral polysaccharide fraction, except that the neutral polysaccharide fraction is used instead of the intermediate polysaccharide fraction to produce the purified neutral polysaccharide fraction. In one embodiment, the neutral polysaccharide fraction is further purified by repeating the aforementioned three steps once. In one embodiment, the neutral polysaccharide fraction is further purified by repeating the aforementioned three steps at least once.

[0147] Those skilled in the art will understand that the neutral polysaccharide fraction can be separated from the first supernatant using any method known in the art. In one embodiment, the separation is performed by filtration, decantation, or centrifugation. In another embodiment, the separation is performed by centrifugation.

[0148] In one embodiment, the neutral polysaccharide grade is separated into solids, oils, waxes, or gums. In one embodiment, the neutral polysaccharide grade is separated into solids. In one embodiment, the neutral polysaccharide grade is separated into solids, and the solid is a powder.

[0149] In one embodiment, after purifying the resulting neutral polysaccharide fraction by repeating the first three steps described above once, the method provides the neutral polysaccharide fraction with a purity of about 80% to about 100%, about 85% to about 100%, about 90% to about 100%, about 95% to about 100%, about 95% to about 98%, or about 95% (w / w) or more.

[0150] In one embodiment, after purifying the neutral polysaccharide fraction once by repeating the first three steps described above, the method provides the neutral polysaccharide fraction in an overall yield of about 10% to about 15% when the intermediate polysaccharide fraction is used as the initial starting material.

[0151] This application includes a method for preparing a neutral polysaccharide fraction (PBG-005), the method comprising:

[0152] Provide intermediate polysaccharide fractions produced by the above method;

[0153] The intermediate polysaccharide fraction is dissolved in water to produce a solution of the intermediate polysaccharide fraction, wherein the solution of the intermediate polysaccharide fraction is from about 1% (w / w) to about 10% (w / w);

[0154] Ethanol is added to the solution of the intermediate polysaccharide fraction to produce a neutral polysaccharide fraction and a first supernatant, wherein the ratio of ethanol to the solution of the intermediate polysaccharide fraction is about 1 to about 1 (v / v) to about 0.5 to about 1 (v / v).

[0155] The neutral polysaccharide fraction is separated from the first supernatant to produce the neutral polysaccharide fraction; and

[0156] Optionally, the neutral polysaccharide fraction can be purified by repeating the preceding three steps.

[0157] This application includes a method for preparing a neutral polysaccharide fraction (PGB-005) from the root of the American ginseng (Panax quinquefolius) plant, the method comprising:

[0158] At temperatures ranging from approximately 10°C to approximately 90°C, using a solution containing approximately 50% (v / v) to approximately 95% (v / v) C 1-3 A solvent mixture of alkyl OH and water is used to extract the root portion from the root portion of Panax quinquefolius to produce a solvent mixture fraction and a first residue, wherein the ratio of the solvent mixture to the root portion is about 20 to about 1 (v / w) to about 1 to about 1 (v / w).

[0159] The first residue is fractionated from the solvent mixture;

[0160] The first residue is extracted with water at a temperature of about 10°C to about 100°C to produce an aqueous fraction containing the intermediate polysaccharide fraction and a second residue, wherein the ratio of water to the root fraction is about 20 to about 1 (v / w) to about 1 to about 1 (v / w).

[0161] The aqueous fraction containing the intermediate polysaccharide fraction is separated from the second residue;

[0162] The aqueous fraction is dried or concentrated to produce the intermediate polysaccharide fraction;

[0163] The intermediate polysaccharide fraction is dissolved in water to produce a solution of the intermediate polysaccharide fraction, wherein the solution of the intermediate polysaccharide fraction is from about 1% (w / w) to about 10% (w / w);

[0164] C 1-3 Alkyl OH groups are added to the solution of the intermediate polysaccharide fraction to produce a neutral polysaccharide fraction and a first supernatant, wherein the C 1-3 The ratio of the alkyl OH to the intermediate polysaccharide fraction solution is about 1 to about 1 (v / v) to about 0.5 to about 1 (v / v);

[0165] The neutral polysaccharide fraction is separated from the first supernatant to produce the neutral polysaccharide fraction; and

[0166] Alternatively, in addition to using the neutral polysaccharide fraction, the neutral polysaccharide fraction can be further purified by repeating the preceding three steps.

[0167] The applicant also obtained an acid-rich polysaccharide fraction (PGB-003) from the intermediate polysaccharide. Therefore, the method further includes preparing the acid-rich polysaccharide fraction. Therefore, the method further includes:

[0168] C 1-3 Alkyl OH is added to the first supernatant to produce a second supernatant rich in acidic polysaccharide fractions, wherein C 1-3 The ratio of alkyl OH to the first supernatant is about 0.5 to about 1 (v / v) to about 8 to about 1 (v / v); and

[0169] The acid-rich polysaccharide fraction is separated from the second supernatant to produce the acid-rich polysaccharide fraction.

[0170] Therefore, this application also includes a method for preparing acid-rich polysaccharide fractions, the method comprising:

[0171] Provide intermediate polysaccharide fractions produced by the above method;

[0172] The intermediate polysaccharide fraction is dissolved in water to produce a solution of the intermediate polysaccharide fraction, wherein the solution of the intermediate polysaccharide fraction is from about 1% (w / w) to about 10% (w / w);

[0173] C 1-3 Alkyl OH groups are added to the solution of the intermediate polysaccharide fraction to produce a neutral polysaccharide fraction and a first supernatant, wherein the C 1-3 The ratio of the alkyl OH to the intermediate polysaccharide fraction solution is about 1 to about 1 (v / v) to about 0.5 to about 1 (v / v);

[0174] The first supernatant was separated from the neutral polysaccharide;

[0175] C 1-3 Alkyl OH is added to the first supernatant to produce a second supernatant rich in acidic polysaccharide fractions, wherein C 1-3 The ratio of alkyl OH to the first supernatant is about 0.5 to about 1 (v / v) to about 8 to about 1 (v / v); and

[0176] The acid-rich polysaccharide fraction is separated from the second supernatant to produce the acid-rich polysaccharide fraction.

[0177] In one embodiment, the intermediate polysaccharide fraction solution is about 2% (w / w) to about 10% (w / w), about 3% (w / w) to about 10% (w / w), about 4% (w / w) to about 10% (w / w), about 5% (w / w) to about 10% (w / w), about 4% (w / w) to about 9% (w / w), about 4% (w / w) to about 8% (w / w), about 4% (w / w) to about 7% (w / w), about 4% (w / w) to about 6% (w / w); about 2% (w / w), about 3% (w / w), about 4% (w / w), about 5% (w / w), or about 6% (w / w). In one embodiment, the intermediate polysaccharide fraction solution is about 5% (w / w).

[0178] In one embodiment, C 1-3 The ratio of the alkyl OH to the intermediate polysaccharide fraction solution is about 1 to about 1 (v / v) to about 0.5 to about 1 (v / v). In one embodiment, C 1-3 The ratio of the alkyl OH to the intermediate polysaccharide fraction solution is about 1 to about 1 (v / v) to about 0.5 to about 1 (v / v) or about 0.7 to about 1 (v / v) to about 0.5 to about 1 (v / v). In one embodiment, when C 1-3 In the step of adding alkyl OH to the solution of the intermediate polysaccharide fraction, C 1-3 The ratio of the alkyl OH to the intermediate polysaccharide fraction solution is about 0.5 to about 1 (v / v).

[0179] In one embodiment, the step of generating the neutral polysaccharide-rich fraction and the first supernatant is performed for more than about 1 hour, more than about 2 hours, more than about 3 hours, more than about 4 hours, more than about 5 hours, or more than about 6 hours. In one embodiment, the step of generating the neutral polysaccharide-rich fraction and the first supernatant is performed for more than about 3 hours. In one embodiment, when the C 1-3 After the alkyl OH is added to the solution of the intermediate polysaccharide fraction, the step of producing the neutral polysaccharide-rich fraction and the first supernatant is carried out for about 3 hours.

[0180] In one embodiment, the C 1-3 The C-alkyl OH groups are present in a ratio of about 0.5 to about 1 (v / v) to about 6 to about 1 (v / v). 1-3 The alkyl OH group is added to the first supernatant at a ratio equal to that of the first supernatant. In one embodiment, the C... 1-3 The C-alkyl OH in a ratio of about 1:1 (v / v) to about 4:1 (v / v) 1-3 The alkyl OH group is added to the first supernatant at a ratio equal to that of the first supernatant. In one embodiment, the C... 1-3 The C-alkyl OH in a ratio of about 2 to about 1 (v / v)1-3 The alkyl OH group was added to the first supernatant at a ratio equal to that of the first supernatant.

[0181] Those skilled in the art will understand that the acid-rich polysaccharide fraction can be separated from the second supernatant by any separation method known in the art. In one embodiment, the aqueous fraction is separated by filtration, decantation, or centrifugation. In one embodiment, the separation is performed by centrifugation.

[0182] Those skilled in the art should understand that, when the C is... 1-3 After the alkyl OH is added to the first supernatant, a sufficiently long time is allowed to generate the acid-rich polysaccharide fraction and the second supernatant. In one embodiment, the step of generating the acid-rich polysaccharide fraction and the second supernatant is carried out for more than about 1 hour, more than about 2 hours, more than about 3 hours, more than about 4 hours, more than about 5 hours, or more than about 6 hours. In one embodiment, the step of generating the acid-rich polysaccharide fraction and the second supernatant is carried out for more than about 3 hours. In one embodiment, the step of generating the acid-rich polysaccharide fraction and the second supernatant is carried out for about 1 hour to about 5 hours, about 2 hours to about 3 hours, about 2 hours to about 4 hours, about 1 hour to about 4 hours, or about 1 hour to about 3 hours. In one embodiment, the step of generating the acid-rich polysaccharide fraction and the second supernatant is carried out for about 1 hour to about 3 hours. In one embodiment, the step of generating the acid-rich polysaccharide fraction and the second supernatant is carried out for about 3 hours.

[0183] In one embodiment, the acid-rich polysaccharide grade is disposed of as a solid, oil, wax, or gum. In one embodiment, the acid-rich polysaccharide grade is disposed of as a solid. In one embodiment, the acid-rich polysaccharide grade is disposed of as a solid, and the solid is a powder.

[0184] In one embodiment, the C in the above steps 1-3 The alkyl OH group is independently selected from methanol, ethanol, propanol, and isopropanol. In one embodiment, the C in the above steps... 1-3 The alkyl OH group is independently selected from methanol and ethanol. In one embodiment, the C in the above steps... 1-3 The alkyl OH group represents ethanol.

[0185] This application includes a method for preparing acid-rich polysaccharide fractions, the method comprising:

[0186] Provide intermediate polysaccharide fractions produced by the above method;

[0187] The intermediate polysaccharide fraction is dissolved in water to produce a solution of the intermediate polysaccharide fraction, wherein the solution of the intermediate polysaccharide fraction is from about 1% (w / w) to about 10% (w / w);

[0188] Ethanol is added to the solution of the intermediate polysaccharide fraction to produce a neutral polysaccharide fraction and a first supernatant, wherein the ratio of ethanol to the solution rich in the intermediate polysaccharide fraction is about 1 to about 1 (v / v) to about 0.5 to about 1 (v / v).

[0189] The first supernatant was separated from the fraction rich in neutral polysaccharides;

[0190] Ethanol is added to the first supernatant to produce a second supernatant rich in acidic polysaccharide fractions, wherein the ratio of ethanol to the first supernatant is about 0.5 to about 1 (v / v) to about 8 to about 1 (v / v); and

[0191] The acid-rich polysaccharide fraction is separated from the second supernatant to produce the acid-rich polysaccharide fraction.

[0192] This application includes a method for preparing acid-rich polysaccharide fractions from the root of the American ginseng (Panax quinquefolius) plant, the method comprising:

[0193] At temperatures ranging from approximately 10°C to approximately 90°C, using a solution containing approximately 50% (v / v) to approximately 95% (v / v) C 1-3 A solvent mixture of alkyl OH and water is used to extract the root portion from the root portion of Panax quinquefolius to produce a solvent mixture fraction and a first residue, wherein the ratio of the solvent mixture to the root portion is about 20 (v / w) to about 1 (v / w) to about 1 (v / w).

[0194] The first residue is fractionated from the solvent mixture;

[0195] The first residue is extracted with water at a temperature of about 10°C to about 100°C to produce an aqueous fraction containing the intermediate polysaccharide fraction and a second residue, wherein the ratio of water to the root fraction is about 20 to about 1 (v / w) to about 1 to about 1 (v / w).

[0196] The aqueous fraction containing the intermediate polysaccharide fraction is separated from the second residue;

[0197] The aqueous fraction is dried or concentrated to produce the intermediate polysaccharide fraction;

[0198] The intermediate polysaccharide fraction is dissolved in water to produce a solution of the intermediate polysaccharide fraction, wherein the solution of the intermediate polysaccharide fraction is from about 1% (w / w) to about 10% (w / w);

[0199] C 1-3Alkyl OH groups are added to the solution of the intermediate polysaccharide fraction to produce a neutral polysaccharide fraction and a first supernatant, wherein the C 1-3 The ratio of the alkyl OH to the intermediate polysaccharide fraction solution is about 1 to about 1 (v / v) to about 0.5 to about 1 (v / v);

[0200] The first supernatant was separated from the neutral polysaccharide;

[0201] C 1-3 Alkyl OH is added to the first supernatant to produce a second supernatant rich in acidic polysaccharide fractions, wherein C 1-3 The ratio of alkyl OH to the first supernatant is about 0.5 to about 1 (v / v) to about 8 to about 1 (v / v); and

[0202] The acid-rich polysaccharide fraction is separated from the second supernatant to produce the acid-rich polysaccharide fraction.

[0203] The applicant also obtained a purified acidic polysaccharide fraction (PGB-007) from the acid-rich polysaccharide fraction. Therefore, the method further includes the preparation of the acidic polysaccharide fraction. Therefore, the method further includes:

[0204] The acid-rich polysaccharide fraction is dissolved in water to produce a solution rich in acid-rich polysaccharide fraction, wherein the solution rich in acid-rich polysaccharide fraction is from about 1% (w / w) to about 10% (w / w);

[0205] C 1-3 Alkyl OH groups are added to the solution rich in the acidic polysaccharide fraction to produce a precipitate and a third supernatant containing the acidic polysaccharide fraction, wherein C 1-3 The ratio of alkyl OH to the solution rich in acidic polysaccharide fraction is about 1 to about 1 (v / v) to about 0.5 to about 1 (v / v);

[0206] The third supernatant containing the acidic polysaccharide fraction is separated from the precipitate; and

[0207] Optionally, the third supernatant may be dried or concentrated to produce the acidic polysaccharide fraction.

[0208] Therefore, this application also includes a method for preparing acidic polysaccharide fractions (PGB-007), the method comprising:

[0209] Provides acid-rich polysaccharide fractions produced by the above method;

[0210] The acid-rich polysaccharide fraction is dissolved in water to produce a solution rich in acid-rich polysaccharide fraction, wherein the solution rich in acid-rich polysaccharide fraction is from about 1% (w / w) to about 10% (w / w);

[0211] C 1-3 Alkyl OH groups are added to the solution rich in the acidic polysaccharide fraction to produce a precipitate and a third supernatant containing the acidic polysaccharide fraction, wherein C 1-3 The ratio of alkyl OH to the solution rich in acidic polysaccharide fraction is about 1 to about 1 (v / v) to about 0.5 to about 1 (v / v);

[0212] The third supernatant containing the acidic polysaccharide fraction is separated from the precipitate; and

[0213] Optionally, the third supernatant may be dried or concentrated to produce the acidic polysaccharide fraction.

[0214] In one embodiment, the solution rich in acidic polysaccharide fraction is about 2% (w / w) to about 10% (w / w), about 3% (w / w) to about 10% (w / w), about 4% (w / w) to about 10% (w / w), about 5% (w / w) to about 10% (w / w), about 4% (w / w) to about 9% (w / w), about 4% (w / w) to about 8% (w / w), about 4% (w / w) to about 7% (w / w), about 4% (w / w) to about 6% (w / w); about 2% (w / w), about 3% (w / w), about 4% (w / w), about 5% (w / w), or about 6% (w / w). In one embodiment, the solution of intermediate polysaccharide fraction is about 5% (w / w).

[0215] In one embodiment, C 1-3 The ratio of the alkyl OH to the intermediate polysaccharide fraction solution is about 1 to about 1 (v / v) to about 0.5 to about 1 (v / v). In one embodiment, C 1-3 The ratio of the alkyl OH to the intermediate polysaccharide fraction solution is about 1 to about 1 (v / v) to about 0.5 (v / v) to about 1 (v / v) or about 0.7 to about 1 (v / v) to about 0.5 to about 1 (v / v) of the C 1-3 Alkyl OH and the solution rich in acidic polysaccharide fractions. In one embodiment, the C 1-3 The ratio of alkyl OH to the solution rich in acidic polysaccharide fraction is about 0.5 to about 1 (v / v).

[0216] In one embodiment, the C 1-3 The alkyl OH group is methanol, ethanol, propanol, or isopropanol. In one embodiment, the C 1-3 The alkyl OH is methanol or ethanol. In one embodiment, the C 1-3 The alkyl OH group represents ethanol.

[0217] In one embodiment, the steps of generating the precipitate and the third supernatant are performed for more than about 1 hour, more than about 2 hours, more than about 3 hours, more than about 4 hours, more than about 5 hours, or more than about 6 hours. In one embodiment, the steps of generating the precipitate and the third supernatant are performed for more than about 1 hour, more than about 2 hours, or more than about 3 hours. In one embodiment, the steps of generating the precipitate and the third supernatant are performed for about 1 hour to about 5 hours, about 2 hours to about 3 hours, about 2 hours to about 4 hours, about 1 hour to about 4 hours, or about 1 hour to about 3 hours. In one embodiment, the steps of generating the precipitate and the third supernatant are performed for about 1 hour to about 3 hours. In one embodiment, the steps of generating the precipitate and the third supernatant are performed for about 3 hours.

[0218] Those skilled in the art will understand that each of the above separation steps can be performed using any method known in the art. In one embodiment, separation is performed by filtration, decantation, or centrifugation. In one embodiment, separation is performed by centrifugation.

[0219] Those skilled in the art will understand that "optionally," for example, "optionally, the third supernatant is dried or concentrated to produce the acidic polysaccharide fraction (PGB-007)," means that the third supernatant containing the acidic polysaccharide fraction can be used in the form produced, or that the solvent from the third supernatant can be removed or concentrated to produce the acidic polysaccharide fraction.

[0220] In one embodiment, the drying or concentration is performed by any solvent removal method known in the art. In one embodiment, the drying or concentration is performed by evaporation. In one embodiment, the evaporation is performed by distillation, open-air evaporation, vacuum evaporation, or rotary evaporation. In one embodiment, the evaporation is performed by rotary evaporation.

[0221] In one embodiment, the drying comprises concentrating the third supernatant containing the acidic polysaccharide fraction, followed by freeze-drying or spray drying to produce the acidic polysaccharide fraction. In one embodiment, the drying comprises concentrating the aqueous fraction containing the acidic polysaccharide fraction, followed by freeze-drying to produce the acidic polysaccharide fraction.

[0222] In one embodiment, the acid-rich polysaccharide fraction produced by the above method can be used without further purification. In another embodiment, as described above, the acid-rich polysaccharide fraction is further purified to produce a purified acid-rich polysaccharide fraction.

[0223] In one embodiment, the acidic polysaccharide grade produced after drying is separated into solid, oil, wax, or gum. In one embodiment, the acidic polysaccharide grade is separated into solid. In another embodiment, the acidic polysaccharide grade is separated into solid, and the solid is a powder.

[0224] In one embodiment, the method provides the acidic polysaccharide fraction with a purity of about 80% (w / w) to about 100% (w / w), about 85% (w / w) to about 100% (w / w), about 90% (w / w) to about 100% (w / w), about 95% (w / w) to about 100% (w / w), about 95% (w / w) to about 98% (w / w), or about 85% (w / w) or more.

[0225] In one embodiment, when the intermediate polysaccharide fraction is used as the initial starting material, the method provides the acidic polysaccharide fraction in an overall yield of about 10% to about 15%.

[0226] This application includes a method for preparing acidic polysaccharide fractions, the method comprising:

[0227] Provides acid-rich polysaccharide fractions produced by the above method;

[0228] The acid-rich polysaccharide fraction is dissolved in water to produce a solution rich in acid-rich polysaccharide fraction, wherein the solution rich in acid-rich polysaccharide fraction is from about 1% (w / w) to about 10% (w / w);

[0229] Ethanol is added to the solution rich in acidic polysaccharide fraction to produce a precipitate and a third supernatant containing the acidic polysaccharide fraction, wherein the ratio of ethanol to the solution rich in acidic polysaccharide fraction is about 1 to about 1 (v / v) to about 0.5 to about 1 (v / v).

[0230] The third supernatant containing the acidic polysaccharide fraction is separated from the precipitate; and

[0231] Optionally, the third supernatant may be dried or concentrated to produce the acidic polysaccharide fraction.

[0232] Alternatively, the acid-rich polysaccharide fraction can be purified using column chromatography to produce the acidic polysaccharide fraction. Accordingly, the method further includes:

[0233] The acid-rich polysaccharide fraction is fractionated from a chromatographic column containing diethylaminoethyl (DEAE) anion exchange resin to produce an acidic polysaccharide elution fraction, wherein the fractionation includes:

[0234] (i) Dissolve the acid-rich polysaccharide fraction in water to produce a solution rich in acid-rich polysaccharide fraction;

[0235] (ii) Loading the solution rich in acidic polysaccharide fractions onto a chromatographic column containing the anion exchange resin; and

[0236] (iii) Eluting the acid-rich polysaccharide-rich elution fraction from the anion exchange resin using ammonium acetate buffer to produce the acid-rich polysaccharide elution fraction, wherein the elution comprises a first step in which the buffer is about 10 mM to about 100 mM ammonium acetate, and a second step in which the buffer is about 0.5 M to about 1.5 M ammonium acetate; and

[0237] Optionally, the acidic polysaccharide elution fraction can be dried or concentrated to produce the acidic polysaccharide fraction.

[0238] Therefore, this application also includes a method for preparing acidic polysaccharide fractions, the method comprising:

[0239] Provides acid-rich polysaccharide fractions produced by the above method;

[0240] The acid-rich polysaccharide fraction is fractionated from a chromatographic column containing diethylaminoethyl (DEAE) anion exchange resin to produce an acidic polysaccharide elution fraction, wherein the fractionation includes...

[0241] (i) Dissolve the acid-rich polysaccharide fraction in water to produce a solution rich in acid-rich polysaccharide fraction;

[0242] (ii) Loading the solution rich in acidic polysaccharide fractions onto a chromatographic column containing the anion exchange resin; and

[0243] (iii) Eluting the acid-rich polysaccharide-rich elution fraction from the anion exchange resin using ammonium acetate buffer to produce the acid-rich polysaccharide elution fraction, wherein the elution comprises a first step in which the buffer is about 10 mM to about 100 mM ammonium acetate, and a second step in which the buffer is about 0.5 M to about 1.5 M ammonium acetate; and

[0244] Optionally, the acidic polysaccharide elution fraction can be dried or concentrated to produce the acidic polysaccharide fraction.

[0245] In one embodiment, the chromatographic column is used in conjunction with a high-performance liquid chromatography (HPLC) system. Therefore, in one embodiment, the chromatographic column is an anion-exchange HPLC column. In one embodiment, the (HPLC) system is... 1200 series preparative HPLC system.

[0246] In one embodiment, the resin comprises crosslinked polymethacrylate.

[0247] In one embodiment, the buffer solution in the first elution step is about 10 mM to about 75 mM ammonium acetate, about 10 mM to about 50 mM ammonium acetate, about 10 mM to about 30 mM ammonium acetate, or about 10 mM to about 20 mM ammonium acetate. In one embodiment, the buffer solution in the first elution step is about 20 mM ammonium acetate.

[0248] In one embodiment, the buffer solution in the second elution step is about 0.75 M to about 1.25 M ammonium acetate. In another embodiment, the buffer solution in the second elution step is about 1 M ammonium acetate.

[0249] In one embodiment, the chromatographic column has a bed size of about 150 mm x about 21.5 mm and a bed volume of about 54.5 mL.

[0250] In one embodiment, the chromatographic column comprising anion exchange resin is 13μm silica gel DEAE-5PW column.

[0251] In one embodiment, when using When fractionation was performed using a 13μm silica gel DEAE-5PW column at a flow rate of approximately 3 ml / min and an injection volume of approximately 5 ml, the acidic polysaccharide eluent fractions were fractionated to correspond to retention times of approximately 42 minutes to approximately 100 minutes.

[0252] The methods for performing anion exchange chromatography are known to those skilled in the art, based on the manufacturer's recommendations regarding flow rate, sample volume, and temperature for the procedure.

[0253] Those skilled in the art will understand that "optionally" in the step of drying or concentrating the acidic polysaccharide elution fraction means that the elution fraction containing the acidic polysaccharide fraction can be used in the produced form or that the solvent can be removed from the elution fraction to produce the acidic polysaccharide fraction.

[0254] In one embodiment, the method provides the acidic polysaccharide fraction with a purity of about 80% (w / w) to about 100% (w / w), about 85% (w / w) to about 100% (w / w), about 90% (w / w) to about 100% (w / w), about 95% (w / w) to about 100% (w / w), about 95% (w / w) to about 98% (w / w), or about 95% (w / w) or more.

[0255] In one embodiment, when the intermediate polysaccharide fraction is used as the initial starting material, the method provides an overall yield of the acidic polysaccharide fraction of about 10% to about 15%.

[0256] In one embodiment, the drying or concentration step is performed by evaporation. In one embodiment, the evaporation is performed by distillation, open-air evaporation, vacuum evaporation, or rotary evaporation. In one embodiment, the evaporation is performed by rotary evaporation.

[0257] In one embodiment, the drying step includes concentrating the acidic polysaccharide elution fraction to produce the acidic polysaccharide fraction, followed by freeze-drying or spray drying to produce the polysaccharide fraction. In another embodiment, the drying includes concentrating the acidic polysaccharide elution fraction containing the acidic polysaccharide fraction, followed by freeze-drying to produce the polysaccharide fraction.

[0258] In one embodiment, C is used in each step of the method for preparing the fractions of this application described above. 1-3 The alkyl OH groups may be the same or different.

[0259] 3. Extraction and fractionation in this application

[0260] The applicant obtained intermediate polysaccharide fractions, purified neutral polysaccharide fractions (PBG-005), acid-rich polysaccharide fractions, and acidic polysaccharide fractions from the root portion of the Panax quinquefolius plant.

[0261] Therefore, this application includes the intermediate polysaccharide fraction (PBG-002) produced by the above method.

[0262] In one embodiment, this application includes an intermediate polysaccharide fraction (PBG-002) containing about 0.5 mol% to about 4 mol% rhamnose (Rha), about 8 mol% to about 20 mol% galacturonic acid (GalA), about 60 mol% to about 80 mol% glucose (Glc), about 3 mol% to about 10 mol% galactose (Gal), and about 3 mol% to about 10 mol% arabinose (Ara). In another embodiment, this application includes an intermediate polysaccharide fraction (PBG-002) containing about 0.5 mol% to about 4 mol% rhamnose (Rha), about 13 mol% to about 17 mol% galacturonic acid (GalA), about 70 mol% to about 74 mol% glucose (Glc), about 3 mol% to about 7 mol% galactose (Gal), and about 3 mol% to about 7 mol% arabinose (Ara).

[0263] In one embodiment, the present application includes an intermediate polysaccharide fraction containing about 1 mol% to about 2 mol% rhamnose (Rha), about 14 mol% to about 16 mol% galacturonic acid (GalA), about 71 mol% to about 73 mol% glucose (Glc), about 4 mol% to about 6 mol% galactose (Gal) and about 4 mol% to about 6 mol% arabinose (Ara).

[0264] In one embodiment, the present application includes an intermediate polysaccharide fraction containing about 0.9 mol% to about 1.7 mol% rhamnose (Rha), about 14.6 mol% to about 15.5 mol% galacturonic acid (GalA), about 72.0 mol% to about 73.0 mol% glucose (Glc), about 4.9 mol% to about 5.7 mol% galactose (Gal) and about 5.0 mol% to about 5.6 mol% arabinose (Ara).

[0265] In one embodiment, the present application includes an intermediate polysaccharide fraction containing about 1.3 mol% rhamnose (Rha), about 15.1 mol% galacturonic acid (GalA), about 72.4 mol% glucose (Glc), about 5.3 mol% galactose (Gal) and about 5.6 mol% arabinose (Ara).

[0266] In one embodiment, this application also includes a neutral polysaccharide fraction (PGB-005) produced by the above method.

[0267] This application also includes a neutral polysaccharide fraction containing about 0.1 mol% to about 5 mol% rhamnose (Rha), about 2 mol% to about 10 mol% galacturonic acid (GalA), about 80 mol% to about 95 mol% glucose (Glc), about 1 mol% to about 5 mol% galactose (Gal), and about 1 mol% to about 5 mol% arabinose (Ara). This application also includes a neutral polysaccharide fraction containing about 0.1 mol% to about 3 mol% rhamnose (Rha), about 3 mol% to about 7 mol% galacturonic acid (GalA), about 87 mol% to about 92 mol% glucose (Glc), about 1 mol% to about 4 mol% galactose (Gal), and about 1 mol% to about 5 mol% arabinose (Ara).

[0268] This application also includes a neutral polysaccharide fraction containing about 0.1 mol% to about 2 mol% rhamnose (Rha), about 5.0 mol% to about 6.0 mol% galacturonic acid (GalA), about 89 mol% to about 92 mol% glucose (Glc), about 1 mol% to about 3 mol% galactose (Gal) and about 1 mol% to about 3 mol% arabinose (Ara).

[0269] This application also includes a neutral polysaccharide fraction containing about 0.1 mol% to about 1.1 mol% rhamnose (Rha), about 5.0 mol% to about 6.0 mol% galacturonic acid (GalA), about 89.3 mol% to about 91.2 mol% glucose (Glc), about 1.4 mol% to about 2.3 mol% galactose (Gal) and about 1.6 mol% to about 2.4 mol% arabinose (Ara).

[0270] This application also includes a neutral polysaccharide fraction containing about 0.6 mol% rhamnose (Rha), about 5.6 mol% galacturonic acid (GalA), about 89.8 mol% glucose (Glc), about 1.9 mol% galactose (Gal) and about 2.0 mol% arabinose (Ara).

[0271] This application also includes a neutral polysaccharide fraction comprising a terminally linked arabinofuranyl residue (t-Araf), a terminally linked glucopyranoyl residue (t-Glcp), a 4-linked mannopyranoyl residue (4-Manp), a 4-linked glucopyranoyl residue (4-Glcp), and a 4,6-linked glucopyranoyl residue (4,6-Glcp).

[0272] This application also includes a neutral polysaccharide fraction comprising about 3% to about 7% terminally linked arabinofuranyl residues (t-Araf), about 3% to about 7% terminally linked glucopyranoyl residues (t-Glcp), about 0.1% to about 4% 4-linked mannopyranoyl residues (4-Manp), about 90% to about 94% 4-linked glucopyranoyl residues (4-Glcp), and about 0.1% to 3% 4,6-linked glucopyranoyl residues (4,6-Glcp).

[0273] This application also includes a neutral polysaccharide fraction comprising about 4.5 wt% to about 5.5 wt% of terminally linked arabinofuranyl residues (t-Araf), about 4.9 wt% to about 5.9 wt% of terminally linked glucopyranoyl residues (t-Glcp), about 0.2 wt% to about 1.2 wt% of 4-linked mannopyranoyl residues (4-Manp), about 91.1 wt% to about 92.1 wt% of 4-linked glucopyranoyl residues (4-Glcp), and about 1.1 wt% to 2.1 wt% of 4,6-linked glucopyranoyl residues (4,6-Glcp).

[0274] This application also includes a neutral polysaccharide fraction comprising about 5.0 wt% terminally linked arabinofuranyl residues (t-Araf), about 5.4 wt% terminally linked glucopyranoyl residues (t-Glcp), about 0.8 wt% 4-linked mannopyranoyl residues (4-Manp), about 91.6 wt% 4-linked glucopyranoyl residues (4-Glcp), and about 1.6 wt% 4,6-linked glucopyranoyl residues (4,6-Glcp).

[0275] In one embodiment, this application includes an acid-rich polysaccharide fraction (PGB-003) produced by the method described above.

[0276] In one embodiment, this application includes an acidic polysaccharide fraction (PGB-007) produced by the method described above.

[0277] This application includes acidic polysaccharide fractions containing carbohydrates of about 1 mol% to about 8 mol% rhamnose (Rha), about 40 mol% to about 50 mol% galacturonic acid (GalA), about 8 mol% to about 16 mol% glucose (Glc), about 14 mol% to about 25 mol% galactose (Gal), and about 15 mol% to about 25 mol% arabinose (Ara). This application also includes acidic polysaccharide fractions containing carbohydrates of about 2 mol% to about 6 mol% rhamnose (Rha), about 43 mol% to about 47 mol% galacturonic acid (GalA), about 10 mol% to about 14 mol% glucose (Glc), about 18 mol% to about 22 mol% galactose (Gal), and about 16 mol% to about 20 mol% arabinose (Ara).

[0278] This application includes an acidic polysaccharide fraction containing about 3 mol% to about 5 mol% rhamnose (Rha), about 45 mol% to about 47 mol% galacturonic acid (GalA), about 11 mol% to about 13 mol% glucose (Glc), about 19 mol% to about 21 mol% galactose (Gal) and about 17 mol% to about 19 mol% arabinose (Ara).

[0279] This application includes an acidic polysaccharide fraction containing about 3.6 mol% to about 4.4 mol% rhamnose (Rha), about 45.1 mol% to about 46.0 mol% galacturonic acid (GalA), about 11.8 mol% to about 12.6 mol% glucose (Glc), about 19.2 mol% to about 20.3 mol% galactose (Gal) and about 17.7 mol% to about 18.7 mol% arabinose (Ara).

[0280] This application also includes an acidic polysaccharide fraction containing about 4.0 mol% rhamnose (Rha), about 45.6 mol% galacturonic acid (GalA), about 12.2 mol% glucose (Glc), about 19.8 mol% galactose (Gal) and about 18.2 mol% arabinose (Ara).

[0281] This application also includes acidic polysaccharide fractions comprising terminally linked rhamnopyranosyl residues (t-Rha), terminally linked arabinose-furanosyl residues (t-Araf), 2-linked rhamnopyranosyl residues (2-Rha), terminally linked glucopyranosyl residues (t-Glcp), terminally linked galactopyranosyl residues and terminally linked galacturonic acid-pyranosyl residues (t-Galp and t-GalA), 4-linked arabinopyranosyl residues or 5-linked arabinose-furanosyl residues (4... (-Arap or 5-Araf), 2,4-linked rhamnopyranosyl residues (2,4-Rha), 3-linked galactopyranosyl residues (3-Galp), 4-linked galactopyranosyl residues and 4-linked galacturonic acid pyranosyl residues (4-Galp and 4-GalA), 4-linked glucopyranosyl residues (4-Glcp), 2,4-linked galactopyranosyl residues (2,4-Galp), and 3,6-linked galactopyranosyl residues (3,6-Galp).

[0282] This application also includes, as an acidic polysaccharide fraction, a component comprising about 2% to about 5% terminally linked rhamnopyranosyl residues (t-Rha), about 3% to about 6% terminally linked arabinose-furanosyl residues (t-Araf), about 1% to about 4% 2-linked rhamnopyranosyl residues (2-Rha), about 3% to about 6% terminally linked glucopyranosyl residues (t-Glcp), about 12% to about 15% terminally linked galactopyranosyl residues and terminally linked galacturonic acid-pyranosyl residues (t-Galp and t-GalA), and about 4% to about 8% 4-linked arabinopyranosyl residues or 5-linked arabinose-furanosyl residues (4-A). The residues are: rap or 5-Araf), about 1% to about 4% 2,4-linked rhamnopyranosyl residues (2,4-Rha), about 1% to about 4% 3-linked galactopyranosyl residues (3-Galp), about 31% to about 35% 4-linked galactopyranosyl residues and 4-linked galacturonic acid pyranosyl residues (4-Galp and 4-GalA), about 22% to about 26% 4-linked glucopyranosyl residues (4-Glcp), about 0.5% to about 3% 2,4-linked galactopyranosyl residues (2,4-Galp), and about 0.5% to about 3% 3,6-linked galactopyranosyl residues (3,6-Galp).

[0283] This application also includes, as an acidic polysaccharide fraction, approximately 3.3% to 4.2% by weight of terminally linked rhamnopyranosyl residues (t-Rha), approximately 4.1% to 5.0% by weight of terminally linked arabinose-furanosyl residues (t-Araf), approximately 2.1% to 3.1% by weight of 2-linked rhamnopyranosyl residues (2-Rha), approximately 4.4% to 5.4% by weight of terminally linked glucopyranosyl residues (t-Glcp), approximately 13.2% to 14.2% by weight of terminally linked galactopyranosyl residues and terminally linked galacturonic acid-pyranosyl residues (t-Galp and t-GalA), and approximately 6.0% to 7.0% by weight of 4-linked arabinopyranosyl residues or 5-linked arabinose-furanosyl residues (4-Rha). -Arap or 5-Araf), about 1.9 wt% to about 2.9 wt% 2,4-linked rhamnopyranosyl residues (2,4-Rha), about 2.0 wt% to about 3.0 wt% 3-linked galactopyranosyl residues (3-Galp), about 32.5 wt% to about 33.3 wt% 4-linked galactopyranosyl residues and 4-linked galacturonic acid pyranosyl residues (4-Galp and 4-GalA), about 23.6 wt% to about 24.4 wt% 4-linked glucopyranosyl residues (4-Glcp), about 0.8 wt% to about 1.8 wt% 2,4-linked galactopyranosyl residues (2,4-Galp), and about 0.8 wt% to about 1.6 wt% 3,6-linked galactopyranosyl residues (3,6-Galp).

[0284] This application also includes, as an acidic polysaccharide fraction, approximately 3.7 wt% of terminally linked rhamnopyranosyl residues (t-Rha), approximately 4.5 wt% of terminally linked arabinose-furanosyl residues (t-Araf), approximately 2.6 wt% of 2-linked rhamnopyranosyl residues (2-Rha), approximately 4.9 wt% of terminally linked glucopyranosyl residues (t-Glcp), approximately 13.7 wt% of terminally linked galactopyranosyl residues and terminally linked galacturonic acid-pyranosyl residues (t-Galp and t-GalA), and approximately 6.4 wt% of 4-linked arabinopyranosyl residues or 5-linked arabinose-furanosyl residues (t-GalA). 4-Arap or 5-Araf), about 2.4 wt% 2,4-linked rhamnopyranosyl residues (2,4-Rha), about 2.5 wt% 3-linked galactopyranosyl residues (3-Galp), about 32.9 wt% 4-linked galactopyranosyl residues and 4-linked galacturonic acid pyranosyl residues (4-Galp and 4-GalA), about 24.0 wt% 4-linked glucopyranosyl residues (4-Glcp), about 1.3 wt% 2,4-linked galactopyranosyl residues (2,4-Galp), and about 1.2 wt% 3,6-linked galactopyranosyl residues (3,6-Galp).

[0285] In one embodiment, the application further includes intermediate polysaccharide fractions, neutral polysaccharide fractions, salts and / or solvates of acid-rich polysaccharide fractions and / or acidic polysaccharide fractions.

[0286] In one embodiment, the salt is an acid addition salt or a base addition salt.

[0287] Those skilled in the art can select appropriate salts (see, for example, SMBerge et al., “Medicinal Salts”, Journal of Pharmaceutical Sciences, 1977, 66, 1-19).

[0288] Acid addition salts suitable for or compatible with the treatment of a subject are any non-toxic organic or inorganic acid addition salts of any basic compound. The basic fractions of this application that form acid addition salts include, for example, fractions of this application containing amine groups. Exemplary inorganic acids that form suitable salts include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid, as well as acidic metal salts such as sodium monohydrogen phosphate and potassium hydrogen sulfate. Exemplary organic acids that form suitable salts include monocarboxylic, dicarboxylic, and tricarboxylic acids. Examples of such organic acids are, for example, acetic acid, trifluoroacetic acid, propionic acid, glycolic acid, lactic acid, pyruvic acid, malonic acid, succinic acid, glutaric acid, fumaric acid, malic acid, tartaric acid, citric acid, ascorbic acid, maleic acid, hydroxymaleic acid, benzoic acid, hydroxybenzoic acid, phenylacetic acid, cinnamic acid, mandelic acid, salicylic acid, 2-phenoxybenzoic acid, p-toluenesulfonic acid, and other sulfonic acids, such as methanesulfonic acid, ethanesulfonic acid, and 2-hydroxyethanesulfonic acid. In one embodiment, a monoacid salt or a diacid salt is formed, and such salts exist in a hydrated, solvated, or substantially anhydrous form. Generally, acid addition salts are more soluble in water and a wide variety of hydrophilic organic solvents and typically exhibit a higher melting point compared to their free base form. The selection criteria for suitable salts are known to those skilled in the art. Other non-pharmaceuticalally acceptable salts, such as, but not limited to, oxalates, may be used, for example, to separate fractions of this application for laboratory use, or for subsequent conversion into pharmaceutically acceptable acid addition salts.

[0289] A base addition salt suitable for or compatible with the treatment of a subject is any non-toxic organic or inorganic base addition salt of any acidic compound. The acidic fraction of this application forming a base addition salt includes, for example, fractions of this application containing a carboxylic acid group. Exemplary inorganic bases forming suitable salts include lithium hydroxide, sodium, potassium, calcium, magnesium, or barium, and ammonia. Exemplary organic bases forming suitable salts include aliphatic, alicyclic, or aromatic organic amines such as isopropylamine, methylamine, trimethylamine, methylpyridine, diethylamine, triethylamine, tripropylamine, ethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, heptaamine, choline, betaine, ethylenediamine, glucosamine, methylglucosamine, theobromine, purine, piperazine, piperidine, N-ethylpiperidine, polyamine resins, etc. Exemplary organic bases are isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine. Choosing a suitable salt can be useful, for example, to prevent the hydrolysis of ester functional groups (if any) elsewhere in the fraction. Criteria for choosing a suitable salt are known to those skilled in the art.

[0290] In one embodiment, the salt is a base addition salt.

[0291] The desired compound salt is formed using standard techniques. For example, the neutral compound is treated with an acid or base in a suitable solvent, and the formed salt is separated by filtration, extraction, or any other suitable method.

[0292] The solvates of the fractions described in this application include those prepared, for example, with pharmaceutically acceptable solvents. Examples of such solvents include water (the resulting solvate is called a hydrate) and ethanol. Those skilled in the art can select suitable conditions for forming a particular solvate.

[0293] In embodiments of this application, one or more fractions of this application described herein may have at least one asymmetric center. When fractions of this application have multiple asymmetric centers, they may exist as diastereomers. It should be understood that all such isomers and mixtures thereof in any proportion are covered within the scope of this application. It should also be understood that while the stereochemistry of a compound may be as shown in any given compound listed herein, such compounds may also contain a certain amount (e.g., less than 20%, suitably less than 10%, more suitably less than 5%) of a compound of this application having an alternative stereochemistry. Any optical isomer, such as isolated, purified, or partially purified optical isomers or racemic mixtures thereof, is included within the scope of this application.

[0294] One or more fractions of this application may exist in different polymorphic forms, and any polymorph or mixture thereof contemplated is included in the scope of this application.

[0295] Any tautomerisms formed by the fractionation of this application and their mixtures are included within the scope of this application.

[0296] 4. The composition of this application

[0297] Using one or more carriers, one or more fractions of this application (e.g., intermediate polysaccharide fraction (PBG-002), neutral polysaccharide fraction (PBG-005), acid-rich polysaccharide fraction, and acidic polysaccharide fraction) are suitably formulated into compositions in a conventional manner. Therefore, this application also includes compositions comprising one or more fractions of this application and carriers. One or more fractions of this application are suitably formulated into pharmaceutical compositions for administration to a subject in a biocompatible form suitable for in vivo administration. Therefore, this application also includes pharmaceutical compositions comprising one or more fractions of this application and a pharmaceutically acceptable carrier. In one embodiment, one or more fractions of this application are suitably formulated into a nutritional pharmaceutical composition for administration to a subject in a biocompatible form suitable for in vivo administration. Therefore, this application also includes nutritional pharmaceutical compositions comprising one or more fractions of this application and a carrier. In one embodiment, the carrier is a pharmaceutically acceptable carrier.

[0298] As those skilled in the art will understand, depending on the chosen route of administration, one or more fractions of this application can be administered to a subject in a variety of forms. One or more fractions of this application can be administered, for example, orally, parenterally, sublingually, sublingually, nasally, rectally, via patch, pump, or transdermally, and pharmaceutical compositions are formulated accordingly. Administration can be carried out periodically or continuously via a pump.

[0299] Parenteral administration includes intravenous, intra-arterial, intraperitoneal, subcutaneous, intramuscular, transepithelial, intranasal, intrapulmonary (e.g., by using an aerosol), intrathecal, rectal, and local (including using patches or other transdermal delivery devices) administration modalities. Parenteral administration can be performed by continuous infusion over a selected time period. Routine procedures and ingredients for selecting and preparing suitable compositions are described, for example, in Remington Pharmaceutical Sciences (2000–20th edition) and the United States Pharmacopeia: National Formulary (USP 24NF19), published in 1999.

[0300] One or more fractions of this application can be administered orally, for example, with an inert diluent or with an assimilated edible carrier, or can be encapsulated in hard or soft-shell gelatin capsules, or can be compressed into tablets, or can be mixed directly with food in a diet. For oral therapeutic administration, one or more fractions can be mixed with excipients and used in the form of ingestible tablets, lozenges, tablets, capsules, pills, granules, tablets, chewing gum, powders, syrups, elixirs, rice paper capsules, aqueous solutions, and suspensions. In the case of tablets, carriers used include lactose, corn starch, sodium citrate, and phosphates. Pharmaceutically acceptable excipients include binders (e.g., pregelatinized corn starch, polyvinylpyrrolidone, or hydroxypropyl methylcellulose); fillers (e.g., lactose, microcrystalline cellulose, or calcium phosphate); lubricants (e.g., magnesium stearate, talc, or silica); disintegrants (e.g., potato starch or sodium glycolate); or wetting agents (e.g., sodium lauryl sulfate). Tablets can be coated using methods known in the art. In the case of tablets, capsules, pouches, pills, or granules intended for oral administration, a pH-sensitive enteric coating, such as Eudragits designed to control the release of the active ingredient, may be optionally used. TMOral dosage forms also include modified release formulations, such as immediate-release and timed-release formulations. Examples of modified release formulations include, for example, sustained release (SR), extended release (ER, XR, or XL), delayed release or timed release, controlled release (CR), or continuous release (CR or Contin), for example, in the form of coated tablets, permeable delivery devices, coated capsules, microcapsules, agglomerated particles (e.g., molecular sieve particles), or fine hollow permeable fiber bundles, or chopped hollow permeable fibers, agglomerated or retained in fiber packages. Timed-release compositions, such as liposomes or those containing one or more fractions, can be formulated with differentially degradable coatings (e.g., by microencapsulation, multiple coatings, etc.). Liposome delivery systems include, for example, small monolayer vesicles, large monolayer vesicles, and multilayer vesicles. Liposomes can be formed from various phospholipids, such as cholesterol, stearamine, or phosphatidylcholine. For oral administration in capsule form, available carriers or diluents include lactose and dried corn starch.

[0301] Liquid formulations for oral administration may be in the form of, for example, solutions, syrups, or suspensions, or suitably as dry products, for reconstitution with water or other suitable carriers prior to use. When oral administration of aqueous suspensions and / or emulsions, one or more fractions of this application are suitably suspended or dissolved in an oil phase in combination with an emulsifier and / or suspending agent. If desired, certain sweeteners and / or flavoring agents and / or coloring agents may be added. Such liquid formulations for oral administration can be prepared by conventional methods with pharmaceutically acceptable additives, such as suspending agents (e.g., sorbitol syrup, methylcellulose, or hydrogenated edible fats); emulsifiers (e.g., lecithin or gum arabic); non-aqueous mediators (e.g., almond oil, esters, or ethanol); and preservatives (e.g., methylparaben, propylparaben, or sorbic acid). Available diluents include lactose and high molecular weight polyethylene glycol.

[0302] One or more fractions of this application can also be freeze-dried and the resulting lyophilized products used, for example, in the preparation of injectable products.

[0303] One or more fractions of this application can also be administered parenterally. Solutions of one or more fractions of this application can be prepared in water appropriately mixed with a surfactant such as hydroxypropyl cellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycol, DMSO, and mixtures thereof with or without alcohol, as well as oils. Under normal storage and use conditions, these formulations contain preservatives to prevent microbial growth. Those skilled in the art will know how to prepare suitable formulations. For parenterally administration, sterile solutions of one or more fractions of this application are typically prepared, and the pH of the solution is appropriately adjusted and buffered. For intravenous injection, the total concentration of the solute should be controlled to make the formulation isotonic. For ocular administration, ointments or dropperable liquids can be delivered using ocular delivery systems known in the art, such as applicators or eye drops. Such compositions may include mucus mimics such as hyaluronic acid, chondroitin sulfate, hydroxypropyl methylcellulose, or polyvinyl alcohol, preservatives such as sorbic acid, EDTA, or benzyl chromium chloride, and a commonly used amount of diluent or carrier. For pulmonary administration, a diluent or carrier suitable for forming an aerosol will be selected.

[0304] One or more fractions of this application can be formulated for parenteral administration by injection, including using conventional catheter insertion techniques or infusion. The formulation for injection can be present in unit dosage forms, such as in ampoules or multi-dose containers, with added preservatives. The composition can take the form of a sterile suspension, solution, or emulsion in an oily or aqueous medium, and may contain formulations such as suspending agents, stabilizers, and / or dispersants. In all cases, the form must be sterile and must be fluid enough for easy injection. Alternatively, one or more fractions of this application may suitably be in sterile powder form for reconstitution with a suitable medium, such as sterile pyrogen-free water, prior to use.

[0305] Compositions for nasal administration can be conveniently formulated into aerosols, drops, gels, and powders.

[0306] For intranasal or inhalation administration, one or more fractions of this application are conveniently delivered in the form of a solution, dry powder formulation, or suspension from a pump-operated spray container squeezed or pumped by a patient, or in the form of an aerosol spray from a pressurized container or nebulizer. Aerosol formulations typically comprise a solution or fine suspension of the active substance in a physiologically acceptable aqueous or non-aqueous solvent and are typically present in a sterile, sealed container in a single- or multiple-dose quantity, which may be used with a nebulizer in cartridge or refill form. Alternatively, the sealed container may be an integral dispensing device, such as a single-dose nasal inhaler or a spray with a metering valve, designed for disposal after use. When the dosage form includes an aerosol, it will contain a propellant, which may be a compressed gas such as compressed air or an organic propellant such as a chlorofluorocarbon. Suitable propellants include, but are not limited to, dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, heptafluoroalkane, carbon dioxide, or other suitable gases. In the case of pressurized aerosols, the dosage unit is appropriately determined by providing a valve to deliver the metered amount. Pressurized containers or nebulizers may contain solutions or suspensions of the active compound. Capsules and cartridges for inhalers or blowpipes (e.g., made of gelatin) may be formulated as powder mixtures containing the compound of this application and a suitable powder matrix (e.g., lactose or starch). Aerosol dosage forms may also be in the form of pump nebulizers.

[0307] Compositions suitable for oral or sublingual administration include tablets, lozenges, and soft lozenges, wherein the active ingredient is formulated with a carrier such as sugar, gum arabic, astragalus gum, or gelatin and glycerin. Compositions for rectal administration are conveniently in the form of suppositories containing a conventional suppository base such as cocoa butter.

[0308] The suppository forms of one or more fractions of this application are suitable for vaginal, urethral, ​​and rectal administration. Such suppositories typically consist of a mixture of substances that are solid at room temperature but melt at body temperature. Substances commonly used to manufacture such media include, but are not limited to, cocoa butter (also known as cocoa brittle), glycerin gelatin, other glycerides, hydrogenated vegetable oils, polyethylene glycol of various molecular weights, and mixtures of fatty acid esters of polyethylene glycol. For further discussion of suppository dosage forms, see, for example: Remington Pharmaceutical Sciences, 16th ed., Mack Publishers, Easton, PA, 1980, pp. 1530-1533.

[0309] One or more fractions of this application may also be coupled to a soluble polymer that serves as a targeted drug carrier. Such polymers may include polyvinylpyrrolidone, pyran copolymers, polyhydroxypropyl methacrylamide-phenol, polyhydroxy-ethyl asparagine-phenol, or polyethylene oxide-polylysine substituted with palmitoyl residues. Furthermore, fractions of this application may be coupled to a class of biodegradable polymers for controlled drug release, such as polylactic acid, polyglycolic acid, copolymers of polylactic acid and polyglycolic acid, poly(ε-caprolactone), polyhydroxybutyric acid, polyorthoesters, polyacetals, polydihydropyran, polycyanoacrylates, and crosslinked or amphiphilic block copolymers of hydrogels.

[0310] In one embodiment, one or more components of this application may be conjugated to a virus, non-virus, or other vector. Viral vectors may include retroviruses, lentiviruses, adenoviruses, herpesviruses, poxviruses, alphaviruses, vaccinia viruses, or adeno-associated viruses. Non-viral vectors may include nanoparticles, cationic lipids, cationic polymers, metal nanoparticles, nanorods, liposomes, micelles, microvesicles, cell-penetrating peptides, or lipid spheres. Nanoparticles may include silica, lipids, carbohydrates, or other pharmaceutically acceptable polymers.

[0311] One or more fractions of this application, including their pharmaceutically acceptable salts and solvates, are suitable for use alone but are generally administered in the form of a pharmaceutical composition in which one or more fractions of this application (the active ingredient) are bound to a pharmaceutically acceptable carrier. Depending on the route of administration, the pharmaceutical composition will comprise about 0.05% to about 99% by weight or about 0.10% to about 70% by weight of the active ingredient (one or more fractions of this application) and about 1% to about 99.95% by weight or about 30% to about 99.90% by weight of a pharmaceutically acceptable carrier, all weight percentages being based on the total composition.

[0312] In one embodiment, one or more fractions of this application are administered simultaneously or sequentially with another therapeutic agent in a separate unit dosage form or together with a single unit dosage form. Therefore, this application provides a single unit dosage form comprising one or more fractions of this application, an additional therapeutic agent, and a pharmaceutically acceptable carrier.

[0313] One or more fractions of this application may be used alone or in combination with another known agent that can be used to treat or prevent viral infections.

[0314] In one embodiment, another known agent that can be used to treat or prevent viral infection is administered or used according to a treatment regimen known to other known agents.

[0315] 4. Methods and uses of this application

[0316] The fraction of this application has been shown to activate Toll-like receptor (TLR) activity and is therefore demonstrated to be a TLR agonist. Consequently, the fraction of this application has been demonstrated to be an immunomodulator, such as an immunostimulant, which activates, for example, the innate and adaptive immune systems, particularly the innate immune system. In in vitro and in vivo models, the fraction of this application has also been shown to provide protection against viral infections. Therefore, the fraction of this application has been demonstrated to have antiviral activity.

[0317] Therefore, this application includes a method for activating Toll-like receptor (TLR) function in cells of a biological sample or a patient, comprising administering an effective amount of one or more fractions of this application to the cells. This application also includes the use of one or more fractions of this application for activating Toll-like receptor (TLR) function in cells, and the use of one or more fractions of this application in the preparation of a medicament for activating Toll-like receptor (TLR) function in cells. This application also includes one or more fractions of this application for activating Toll-like receptor (TLR) function in cells.

[0318] The fractions in this application have also been shown to stimulate the production of chemokines, particularly interleukin-8 (IL-8).

[0319] Therefore, this application also includes a method for stimulating the production of one or more chemokines in cells of a biological sample or a patient, comprising administering an effective amount of one or more fractions of this application to the cells. This application also includes the use of one or more fractions of this application for stimulating the production of one or more chemokines in cells, and the use of one or more fractions of this application in the preparation of a medicament for stimulating the production of one or more chemokines. This application also includes one or more fractions of this application for stimulating the production of one or more chemokines in cells.

[0320] In one embodiment, one or more chemokines are selected from IL-8, IL-2, IL-1, IL-6, INF-γ, TNF-α, and interferon-γ. In one embodiment, one or more chemokines are IL-8.

[0321] Since the fraction of this application has been shown to activate the activity of Toll-like receptor (TLR) functional proteins and stimulate the production of one or more chemokines, the fraction of this application can be used to treat diseases, disorders, or conditions by activating Toll-like receptor (TLR) function or by stimulating the production of one or more chemokines. Therefore, the fraction of this application can be used as a pharmaceutical. In one embodiment, the fraction of this application can be used as a nutritional product or functional food.

[0322] This application also includes a method for treating a disease, disorder, or condition by activating TLR function, the method comprising administering a therapeutically effective amount of one or more fractions of this application to a subject in need of such treatment.

[0323] Therefore, this application also includes the use of one or more fractions of this application to treat diseases, disorders, or conditions by activating TLR function, and the use of one or more fractions of this application to prepare a medicament for treating diseases, disorders, or conditions by activating TLR function. This application also includes one or more fractions of this application for treating diseases, disorders, or conditions by activating TLR function.

[0324] In one embodiment, the TLR is TLR2 or TLR4. In one embodiment, the TLR is a dimer. In one embodiment, the TLR is a homodimer.

[0325] This application also includes a method for treating a disease, disorder, or condition by stimulating the production of one or more chemokines, the method comprising administering a therapeutically effective amount of one or more fractions of this application to a subject in need of such treatment.

[0326] This application also includes the use of one or more fractions of this application for treating diseases, disorders, or conditions by stimulating the production of one or more chemokines, and the use of one or more fractions of this application for preparing a medicament for treating diseases, disorders, or conditions by stimulating the production of one or more chemokines. This application also includes one or more fractions of this application for treating diseases, disorders, or conditions by stimulating the production of one or more chemokines.

[0327] Toll-like receptors (TLRs) are known to be involved in the activation of innate and adaptive immune responses. In particular, they are well known to be responsible for initiating adaptive and innate immune responses.

[0328] Therefore, this application also includes a method for activating innate and / or subsequent adaptive responses, whether in a biological sample or in the cells of a patient, the method comprising administering a therapeutically effective amount of one or more fractions of this application to the cells. This application also includes the use of one or more fractions of this application for activating innate and / or adaptive innate responses in cells, and the use of one or more fractions of this application for preparing a medicament for activating innate and / or adaptive innate responses in cells. This application also includes one or more fractions of this application for activating innate and / or adaptive innate responses in cells.

[0329] This application also includes a method for treating a disease, disorder, or condition associated with the activation of innate and / or adaptive innate responses, the method comprising administering a therapeutically effective amount of one or more fractions of this application to a subject in need of such treatment.

[0330] This application also includes the use of one or more fractions of this application for treating diseases, disorders, or conditions associated with the activation of innate and / or adaptive innate responses, and the use of one or more fractions of this application for preparing medicaments for treating diseases, disorders, or conditions associated with the activation of innate and / or adaptive innate responses. This application also includes one or more fractions of this application for treating diseases, disorders, or conditions associated with the activation of innate and / or adaptive innate responses.

[0331] In one embodiment, a treatable disease, disorder, or condition associated with activation of the TLR function, stimulation of the production of one or more chemokines, and / or activation of innate and / or adaptive innate responses is treated as a viral infection, microbial infection, or cancer. Therefore, this application also includes a method of treating a viral infection, microbial infection, or cancer, the method comprising administering a therapeutically effective amount of one or more fractions of this application to a subject in need of it.

[0332] The fractions in this application have demonstrated antiviral activity against viral infections such as influenza virus infection and herpes simplex virus (HSV) infection, such as HSV-1 and HSV-2.

[0333] Therefore, this application also includes a method for treating or preventing viral infection, the method comprising administering a therapeutically effective amount of one or more fractions of this application to a subject in need thereof.

[0334] This application also includes the use of one or more fractions of this application for treating or preventing viral infections, and the use of one or more fractions of this application for preparing a medicament for treating or preventing viral infections. This application also includes one or more fractions of this application for treating or preventing viral infections.

[0335] In one embodiment, the viral infection is an influenza virus infection, a herpes virus infection, a human immunodeficiency virus (HIV) infection, a respiratory syncytial virus (RSV) infection, a variety of cold and flu viruses, or a coronavirus infection.

[0336] In one embodiment, influenza virus infection is an infection with at least one influenza virus. In one embodiment, the influenza virus is an influenza A virus or an influenza B virus. In one embodiment, the influenza A virus is a hemagglutinin (“H”) or neuraminidase (“N”) subtype. In one embodiment, the influenza A virus subtype is selected from the following types: influenza A virus subtype H1N1, influenza A virus subtype H1N2, influenza A virus subtype H2N2, influenza A virus subtype H2N3, influenza A virus subtype H3N1, influenza A virus subtype H3N2, influenza A virus subtype H3N8, influenza A virus subtype H5N1, influenza A virus subtype H5N2, influenza A virus subtype H5N3, and influenza A virus subtype H5N4. The influenza A virus subtypes are H5N6, H5N8, H5N9, H6N1, H6N2, H7N1, H7N2, H7N3, H7N4, H7N7, H7N9, H9N2, and H10N7. In one embodiment, the influenza A virus subtype is influenza A virus subtype H5N1.

[0337] In one embodiment, the viral infection is an infection with the H1N1 subtype of influenza A virus.

[0338] In one embodiment, the influenza A virus is a variant of the influenza A virus. In one embodiment, the influenza A virus variant originates from avian influenza virus, human influenza virus, swine influenza virus, equine influenza virus, bat influenza virus, feline influenza virus, and canine influenza virus. In one embodiment, the influenza A virus is a human influenza virus.

[0339] In one embodiment, herpesvirus infection is an infection with at least one herpesvirus. In one embodiment, the herpesvirus infection is selected from herpes simplex virus 1 (HSV-1) infection, herpes simplex virus (HSV-2), varicella-zoster virus (VZV) infection, Epstein-Barr virus (EBV) infection, human cytomegalovirus (HCMV) infection, human herpesvirus 6A (HHV-6a) infection, human herpesvirus 6A (HHV-6B) infection, human herpesvirus 7 (HHV-7), human herpesvirus 6A, and Kaposi's sarcoma-associated herpesvirus (KSHV) infection.

[0340] In one embodiment, the herpesvirus infection is selected from herpes simplex virus 1 (HSV-1) infection and herpes simplex virus (HSV-2) infection.

[0341] In one embodiment, human immunodeficiency virus (HIV) infection is either HIV-1 infection or HIV-2 infection.

[0342] In one embodiment, the coronavirus infection is either a severe acute respiratory syndrome coronavirus (SARS)-related coronavirus infection or a Middle East respiratory syndrome-related coronavirus (MERS)-related coronavirus infection. Therefore, in one embodiment, the MERS-related coronavirus infection is MERS-CoV or a MERS-CoV variant infection.

[0343] In one embodiment, SARS-related coronavirus infection is infection with at least one SARS-related coronavirus. In one embodiment, the SARS-related coronavirus is SARS-CoV or SARS-CoV-2. In one embodiment, the SARS-related coronavirus is a variant of the SARS-related coronavirus. In one embodiment, the SARS-related coronavirus variant is a variant of SARS-CoV or a variant of SARS-CoV-2. Therefore, in one embodiment, SARS-related coronavirus infection is SARS-CoV-2 infection or SARS-CoV-2 variant infection. In one embodiment, SARS-related coronavirus infection is SARS-CoV-2 infection.

[0344] In one embodiment, MERS-related coronavirus infection is an infection with at least one MERS-related coronavirus. In one embodiment, the MERS-related coronavirus is MERS-CoV. In one embodiment, the MERS-related coronavirus is a variant of the MERS-related coronavirus. In one embodiment, the MERS-related coronavirus variant is a variant of MERS-CoV.

[0345] "Variants" refer to, for example, viruses that contain one or more changes or mutations in the nucleotide sequence of the original viral genome.

[0346] In one embodiment, the viral infection is either a primary viral infection or an opportunistic infection.

[0347] Those skilled in the art should also understand that a disease, disorder, or symptom caused by a viral infection can be a primary disease, disorder, or symptom caused by a viral infection, or it can be a secondary disease, disorder, or symptom caused by a viral infection, as a result of a primary disease, disorder, or symptom caused by a viral infection.

[0348] In one embodiment, the cells are in vivo. In another embodiment, the cells are in vitro.

[0349] This application also includes a method for treating a disease, disorder, or condition that is treatable by activating TLR function, by stimulating the production of one or more chemokines, and / or associated with activating innate and / or adaptive innate responses, the method comprising administering to a subject in need a therapeutically effective amount of one or more fractions of this application, in combination with another known agent that is treatable by activating TLR function, by stimulating the production of one or more chemokines, and / or associated with activating innate and / or adaptive innate responses. This application also includes the use of one or more fractions of this application in combination with another known pharmaceutical agent, said known pharmaceutical agent being used to treat diseases, disorders, or conditions that are treatable by activating TLR function, by stimulating the production of one or more chemokines, and / or related to the activation of innate and / or adaptive innate responses. The above uses are for treating diseases, disorders, or conditions that are treatable by activating TLR function, by stimulating the production of one or more chemokines, and / or related to the activation of innate and / or adaptive innate responses. The above uses are for preparing a medicament for treating diseases, disorders, or conditions that are treatable by activating TLR function, by stimulating the production of one or more chemokines, and / or related to the activation of innate and / or adaptive innate responses. This application also includes a method for treating diseases, disorders, or conditions that are treatable by activating TLR function, stimulating the production of one or more chemokines, and / or associated with activating innate and / or adaptive innate responses, by combining one or more fractions of this application with another known agent for treating diseases, disorders, or conditions that are treatable by activating TLR function, stimulating the production of one or more chemokines, and / or associated with activating innate and / or adaptive innate responses. In one embodiment, the disease, disorder, or condition that is treatable by activating TLR function, stimulating the production of one or more chemokines, and / or associated with activating innate and / or adaptive innate responses is a viral or microbial infection, such as influenza virus infection, herpes virus infection, human immunodeficiency virus (HIV) infection, respiratory syncytial virus (RSV) infection, or coronavirus infection.

[0350] Those skilled in the art should understand that the subject may simultaneously suffer from two or more diseases, disorders, or conditions caused by viral infection.

[0351] In another embodiment, this application includes a method of treating or preventing a viral infection, the method comprising administering a therapeutically effective amount of one or more fractions of this application to a subject in need. In one embodiment, one or more fractions of this application are administered in combination with another known agent that can be used to treat or prevent a viral infection.

[0352] In one embodiment, known agents for treating or preventing viral infections are selected from antiviral agents, antiretroviral agents, corticosteroids, antimalarial agents, antibiotics, and immunotherapies, and combinations thereof. In one embodiment, an antiviral agent is selected from remdesivir, peramivir, zanamivir, oseltamivir, and balozavir, and combinations thereof. In one embodiment, an antiretroviral agent is selected from lopinavir and ritonavir, and combinations thereof. In one embodiment, a corticosteroid is selected from cortisone, prednisone, and methylprednisolone, and combinations thereof. In one embodiment, an antimalarial agent is chloroquine. In one embodiment, an antibiotic is selected from macrolide antibiotics, fluoroquinolones, tetracycline antibiotics, and combinations thereof. In one embodiment, a macrolide antibiotic is selected from azithromycin and clarithromycin, and combinations thereof. In one embodiment, a fluoroquinolone antibiotic is selected from ciprofloxacin and levofloxacin, and combinations thereof. In one embodiment, a tetracycline is selected from doxycycline and tetracycline, and combinations thereof.

[0353] In one embodiment, the immunotherapy is an antibody therapy or a vaccine. In one embodiment, the antibody therapy is casirivimab, imdevimab, or bamlanivimab, or a combination thereof. In one implementation, the antibody therapy is casirivimab or imdevimab, or a combination thereof. In one embodiment, the antibody therapy is casirivimab and imdevimab. In one embodiment, the antibody therapy is bamlanivimab.

[0354] In one embodiment, the immunotherapy is a vaccine. In one embodiment, the vaccine is a Covid-19 vaccine. In one embodiment, the Covid-19 vaccine is any approved or developing Covid-19 vaccine. In one embodiment, the vaccine is BNT162b2, AZD1222, Ad5-nCoV, mRNA-1273, CoronaVac, Gam-Covid-Vac, Ad26.COV2.S, NVX-CoV237, BBV152, CoVLP, CVnCoV, INO-4800, EpiVacCorona, INO-4800, AG0301-COVID-19, LV-SMENP-DC, LNP-nCoVsaRNA, GX-19, SCB-2019, COVAX-19, Lunar-COV19 / ARCT-021, CoVLPs, COVID-19 / aAPC, SARS-CoV-2Sclamp, GRAd-Cov2, AD5-nCOV, SputnikV, or a combination thereof. In one embodiment, the vaccine is Ad26.COV2.S. In one embodiment, the vaccine is BNT162b2. In one embodiment, the vaccine is AZD1222. In one embodiment, the vaccine is mRNA-1273.

[0355] One or more fractions of this application can enhance the efficacy of vaccines used to treat viral infections. Therefore, one or more fractions of this application can be used as vaccine adjuvants.

[0356] Therefore, this application includes a method for improving the efficacy of a vaccine for treating viral infections, the method comprising administering a therapeutically effective amount of one or more fractions of this application to a subject in need of it.

[0357] This application also includes the use of one or more fractions of this application for improving the efficacy of vaccines for treating viral infections, and the use of one or more fractions of this application in the preparation of a medicament for improving the efficacy of vaccines for treating viral infections. This application also includes one or more fractions of this application for improving the efficacy of vaccines for treating viral infections.

[0358] In one embodiment, one or more fractions of this application are administered simultaneously or sequentially with another agent in a separate unit dosage form or together in a single unit dosage form.

[0359] In one embodiment, one or more fractions of this application are applied or used as soon as possible after exposure to or potential exposure to the virus. In another embodiment, one or more fractions of this application are applied or used until treatment of the viral infection is achieved. For example, until the virus is completely eliminated, or until the viral load is reduced to a level where the subject's defenses are no longer overwhelmed and can kill any remaining virus.

[0360] The effective amount can vary depending on factors such as the subject's disease state, age, sex, and / or weight. The amount of a given compound corresponding to this amount will vary depending on various factors, such as the given drug or compound, the drug formulation, the route of administration, the condition, the type of disease or disorder, the identity of the subject receiving treatment, etc., but can still be routinely determined by those skilled in the art. The effective amount is the amount at which any improvement or reduction of disease symptoms is observed after treatment.

[0361] In one embodiment, one or more fractions of this application may be administered at least once a week. In one embodiment, for a given treatment, one or more fractions may be administered to the subject about once every three weeks, or about once a week to about once daily. In another embodiment, one or more fractions may be administered 2, 3, 4, 5, or 6 times daily. The length of treatment depends on a variety of factors, such as the severity of the disease, disorder, or condition, the age of the subject, the concentration and / or activity of one or more fractions of this application, and / or combinations thereof. It should also be understood that the effective dose of the compound used for treatment may be increased or decreased during a particular treatment regimen. Changes in dose may occur and become apparent by standard diagnostic assays known in the art. In some cases, prolonged dosing may be required. For example, one or more fractions may be administered to the subject in a dose and duration sufficient to treat the patient.

[0362] The fractions of this application can be used alone or in combination with other known agents for treating diseases, disorders, or conditions that are treatable by activating TLR function, by stimulating the production of one or more chemokines, and / or by activating innate and / or adaptive innate responses, such as those associated with viral infections. When used in combination with other agents that are treatable by activating TLR function, by stimulating the production of one or more chemokines, and / or by activating innate and / or adaptive innate responses, in one embodiment, one or more fractions of this application are administered simultaneously with those agents. As used herein, “simultaneous administration” of two substances to a subject means providing each of the two substances such that they are simultaneously biologically active in the individual. Specific details of administration will depend on the pharmacokinetics of the two substances in the presence of each other, and may include administration of the two substances over several hours, or even administration of the other substance within 24 hours of administration of one substance, if the pharmacokinetics are suitable. The design of suitable dosing regimens is conventional to those skilled in the art. In specific embodiments, the two substances will be administered substantially simultaneously, i.e., within minutes of each other, or in a single composition comprising the two substances. Another embodiment of this application is the administration of a combination of pharmaceutical agents to a subject in a non-simultaneous manner. In one embodiment, one or more fractions of this application are administered simultaneously or sequentially with another therapeutic agent in a separate unit dosage form or together with a single unit dosage form. Therefore, this application provides a single unit dosage form comprising one or more fractions of this application, an additional therapeutic agent, and a pharmaceutically acceptable carrier.

[0363] The dosage of one or more fractions of this application can vary depending on many factors, such as the pharmacodynamic properties of the compound, the route of administration, the recipient's age, health status and weight, the nature and severity of symptoms, the frequency of treatment and the type of concurrent treatment (if any), and the clearance rate of the compound in the treated subject. Those skilled in the art can determine an appropriate dosage based on the above factors. The fractions of this application can initially be administered at an appropriate dosage, which can be adjusted as needed based on clinical response. Typically, a dosage is selected to maintain serum levels of the fractions of this application at about 0.01 μg / cc to about 1000 μg / cc, or about 0.1 μg / cc to about 100 μg / cc. As a representative example, for adults, the oral dosage range of one or more fractions of this application is about 1 mg to about 1000 mg daily, suitably about 1 mg to about 500 mg daily, and more suitably about 1 mg to about 200 mg daily. For parenteral administration, representative doses are about 0.001 mg / kg to about 10 mg / kg, about 0.01 mg / kg to about 10 mg / kg, about 0.01 mg / kg to about 1 mg / kg, or about 0.1 mg / kg to about 1 mg / kg. For oral administration, representative doses are about 0.001 mg / kg to about 10 mg / kg, about 0.1 mg / kg to about 10 mg / kg, about 0.01 mg / kg to about 1 mg / kg, or about 0.1 mg / kg to about 1 mg / kg. For suppository administration, representative doses are about 0.1 mg / kg to about 10 mg / kg, or about 0.1 mg / kg to about 1 mg / kg. In one embodiment of this application, the composition is formulated for oral administration, and one or more fractions are suitably in tablet form containing 0.25 mg, 0.5 mg, 0.75 mg, 1.0 mg, 5.0 mg, 10.0 mg, 20.0 mg, 25.0 mg, 30.0 mg, 40.0 mg, 50.0 mg, 60.0 mg, 70.0 mg, 75.0 mg, 80.0 mg, 90.0 mg, 100.0 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, 600 mg, 650 mg, 700 mg, 750 mg, 800 mg, 850 mg, 900 mg, 950 mg, or 1000 mg of active ingredient per tablet. The fractions of this application may be administered in a single dose daily, weekly, or monthly, or the total daily dose may be divided into two, three, or four daily doses.

[0364] In one embodiment, the fractions of this application and / or compositions thereof are used or administered alone in the methods or uses described herein, or in combination with other known agents used in the methods or uses described herein, for example, for the treatment of viral infections.

[0365] In one embodiment, the fractions and / or compositions thereof of this application may be used as additives, for example, in nutritional product compositions. Therefore, this application also includes the use of one or more fractions and / or compositions thereof of this application as additives.

[0366] In one embodiment, the subject is a mammal. In another embodiment, the subject is a human.

[0367] 5. Examples

[0368] The following non-limiting embodiments are used to illustrate this application.

[0369] Example 1: Preparation and analysis of PBG-005 and PBG-007

[0370] Figure 1 A flowchart illustrating an embodiment of a method for separating intermediate polysaccharide fractions (PBG-002) and neutral polysaccharide fractions (PBG-005), as well as acid-rich polysaccharide fractions and acidic polysaccharide fractions (PBG-007), is shown.

[0371] Step 1. Preparation of intermediate polysaccharide fraction PBG-002

[0372] Ground North American ginseng root (20-40 mesh) was refluxed with 85% ethanol at a ratio of 10:1 (V / w) for 3 hours. The extract was separated, and the residue was refluxed with water at a ratio of 12:1 (V / w) for 3 hours. The extract was centrifuged, concentrated, and spray-dried to obtain PBG-002 bulk powder.

[0373] Step 2. Preparation of neutral polysaccharide fraction PBG-005

[0374] Ethanol was added to an aqueous solution of PBG-002 (5%) at a ratio of 0.5:1. The mixture was kept at room temperature for 3 hours. The precipitate was separated by centrifugation. The precipitate was redissolved in water (5%) and the same ethanol precipitation procedure was performed to produce the neutral fraction PBG-005. The yield of PBG-005 prepared from PBG-002 was approximately 10-15%, with a purity greater than 95%.

[0375] Step 3. Preparation of acid polysaccharide fractions PBG-003 and PBG-007.

[0376] Another portion of ethanol was added to the supernatant from the first ethanol precipitation in the preparation of the neutral fraction, with an ethanol-to-supernatant ratio of 2:1 (v / v). The mixture was kept at room temperature for 3 hours. The precipitate was separated by centrifugation and dried to obtain the acid-rich polysaccharide fraction PBG-003. The acid-rich polysaccharide fraction was then dissolved in water (5%), and ethanol was added to the solution at a ratio of 0.5:1. The mixture was stirred and kept at room temperature for 3 hours. The precipitate was separated by centrifugation. The supernatant was concentrated and lyophilized to obtain the dry powder acid polysaccharide PBG-007-1. The yield of the acid polysaccharide fraction PBG-007 (PBG-007-1) from PBG-002 was approximately 10-15%, with a purity greater than 85%.

[0377] Step 4. Chromatographic purification of the acid fraction to produce the acidic polysaccharide fraction PBG-007

[0378] The acid-rich polysaccharide fraction was then further purified by ion-exchange chromatography. First, the starting material PBG-003 was dissolved in Milli-Q water at a concentration of 50 mg / ml, then filtered through a 0.45 μm syringe filter unit before HPLC. The solution was injected into an Agilent 1200 preparative HPLC system equipped with an ELSD detector and a FoxyR2 fraction collector using a multiple injection method (5 injections per round, 5 ml injection volume per injection). Detailed chromatographic conditions are as follows.

[0379] Instrument: Agilent 1200 series preparative HPLC system

[0380] Chromatographic column: TSK silica gel 13μm DEAE-5PW column, 150mm x 21.5mm (54.5ml)

[0381] Detector: Agilent 1200 Series ELSD (60℃, 3.5 bar)

[0382] Elution Buffer A: 20mM NH4OAc Elution Buffer B: 1M NH4OAc

[0383] Flow rate: 3 ml / min; Injection volume: 5 mL

[0384] Runtime: 110 minutes; Processing time: 2 minutes

[0385] The required fractions (retention times from 42 minutes to 100 minutes) were collected and lyophilized to obtain a dry powder as product (PBG-007-2). The yield of product (PBG-007-2) from PBG-002 was approximately 10-15%, with a purity greater than 95%.

[0386] Example 2: Structural characterization of neutral and acidic polysaccharides

[0387] 1. Component sugar analysis

[0388] The component sugar composition of the isolated neutral and acidic polysaccharides was determined by hydrolysis of the polysaccharide chains, releasing their component sugars and further quantifying the released component sugars. Table 1 lists the composition of typical samples from the above process. The composition of crude polysaccharides is also listed for comparison.

[0389] Table 1: Sugar composition of typical acidic and neutral polysaccharides

[0390]

[0391] 2. Glycosyl linkage analysis

[0392] Glycosyl linkage (NaOH method):

[0393] For glycosyl linkage analysis, the sample was fully methylated, depolymerized, reduced, and acetylated; and the resulting partially methylated sugar alcohol acetates (PMAAs) were analyzed by gas chromatography-mass spectrometry (GC-MS), for example, York et al. (1985) Enzymol method, 118:3-40.

[0394] Initially, the analyzed aliquots were suspended in approximately 200 μl of dimethyl sulfoxide (DMSO). The samples were then fully methylated using the method of Ciukanu and Kerek (1984) CarboHydr, reference 131:209-217 (treatment with sodium hydroxide and iodomethane in dry DMSO). The samples were placed in NaOH base for 10 minutes, followed by the addition of iodomethane and incubation for 20 minutes. Base was then added over 10 minutes, and finally, more iodomethane was added over 20 minutes. The addition of more iodomethane and NaOH base ensured complete methylation of the polymer. After sample treatment, the fully methylated material was reduced to methyl uronic acid via hyperdeuterium, followed by hydrolysis with 2M trifluoroacetic acid (hydrolysis at 121°C for 2 hours in a sealed tube), reduction with NaBD4, and acetylation with acetic anhydride / trifluoroacetic acid. The obtained PMAA was analyzed on a Hewlett Packard 5890 GC connected to a 5970 MSD (mass selection detector, electron bombardment ionization mode); and separated on a 30m Supelco 2330 bonded phase fused silica capillary column.

[0395] Glycosyl linkage (Hakamori method)

[0396] For glycosyl linkage analysis, the sample was methylated by an improvement on the Hakomori method; depolymerized, reduced and acetylated; and the resulting partially methylated sugar alcohol acetates (PMAAs) were analyzed by gas chromatography-mass spectrometry (GC-MS), for example York et al. (1985) Enzymol method, 118:3-40.

[0397] In summary, after lyophilization, aliquots were removed from the sample, suspended in approximately 200 μL of dimethyl sulfoxide, and placed on a magnetic stirrer for 5 days. 0.7 mL of potassium dimethylsulfonate (3.6 M) was added. After stirring at room temperature for 8 hours, the reaction mixture was cooled to 0 °C, excess iodomethane (0.7 mL) was added, and the tube was sealed. Incubation continued overnight at room temperature. After sample treatment, the sample was dried under nitrogen, and approximately 200 μL of hyperdeuterium was added. Incubation was performed at room temperature for 1 hour and 45 minutes, followed by bypassing through an on-guard H column. After drying under nitrogen, the sample was again fully methylated using the method of Ciukanu and Kerek (1984) CarboHydr, reference 131:209-217 (treatment with sodium hydroxide and iodomethane in dry DMSO). The sample was placed in NaOH base for 10 minutes, followed by the addition of iodomethane and incubation for 40 minutes. More NaOH base and iodomethane were then added over 40 minutes. Additional iodomethane and NaOH base were added to ensure complete methylation of the polymer. The fully methylated material was hydrolyzed using 2M trifluoroacetic acid (hydrolysis in a sealed tube at 100°C for 2 hours), reduced with NaBD4, and acetylated using acetic anhydride / pyridine. The resulting PMAA was analyzed on a Hewlett Packard 5890 GC connected to a 5970 MSD (mass-selective detector, electron impact ionization mode); separation was performed on a 30m Supelco 2330 bonded-phase fused silica capillary column.

[0398] result

[0399] Tables 2 and 3 summarize the glycosyl linkage information of typical neutral and acidic polysaccharide samples obtained through the above process.

[0400] Table 2: Glycosyl linkages of neutral polysaccharides

[0401]

[0402]

[0403] Table 3: Glycosyl linkages of neutral polysaccharides

[0404]

[0405]

[0406] As shown above, the neutral polysaccharide PBG-005 mainly contains glucose as its structural component, with over 90% being 4-glucose and other minor linkages. The acidic polysaccharide PBG-007 contains galacturonic acid, arabinose, rhamnose, galactose, and glucose as component sugars, with galacturonic acid being the main component. It primarily contains terminal GalA, 4-GalA, 4-Glc, and other minor linkages.

[0407] Example 3: Toll-like receptor activity

[0408] method:

[0409] HEK293 cells were constructed to stably express specific human TLRs (TLR2 or TLR4). In this experiment, HEK293 cells were treated with polysaccharide fractions at concentrations of 100 μg / mL, 500 μg / mL, or 2000 μg / mL for 24 hours. The output of IL-8 stimulated by TLR2 and TLR4 in the supernatant was measured by ELISA.

[0410] Positive controls: Pam3CSK4 for TLR2 and LPS for TLR4.

[0411] In this study, PBG-005 showed mixed activity affecting TLR2 and TLR4, while PBG-007-1 showed minimal TLR2 activity and strong TLR4 activity (see [link to study]). Figure 2 and 3 PBG-002 exhibits higher activity towards TLR2 than towards TLR4. The TLR2 activity of the three exemplary fractions of this application was compared as follows: PBG-005 > PBG-002 > PBG-007-1. Regarding TLR4 activity: PBG-007-1 > PBG-005 > PBG-002.

[0412] Example 4: Effects against H1N1 influenza virus

[0413] Macrophages (in vitro)

[0414] method:

[0415] Primary human macrophages were pretreated for 24 hours with different concentrations of PBG-002, PBG-005, PBG-007-1, or a single culture medium. In the presence of the exemplary fraction of this application, virus was added at 0.1 MOI and the cells were cultured for 40 minutes. The virus was then washed away, and fresh culture medium and the exemplary fraction of this application were added, followed by culturing for 24 hours. The supernatant was collected, and the viral titer was determined by adding a dilution of the supernatant to susceptible cell lines.

[0416] Discussion: The first study showed that PBG-005 and PBG-007-1 had a significant effect on H1N1 titers, although the dose-response curves were unclear, while PBG-002 had no effect on titers (see [link to study]). Figure 4A and 4B A follow-up study at higher doses showed that PBG-005 or PBG-002 had no activity against viral titers, contradicting the results of PBG-005 in earlier replication experiments. PBG-007-1 showed a significant dose-response effect on H1N1 titers, reaching significance only at the two highest doses.

[0417] Example 5: Effects on HSV-1

[0418] a) Mouse dermal cells (in vitro)

[0419] method:

[0420] Primary mouse dermal fibroblasts were pretreated for 24 hours with different concentrations of PBG-002, PBG-005, PBG-007-1, or a single culture medium. In the presence of the exemplary fraction of this application, virus was added at 0.1 MOI and the cells were cultured for 40 minutes. The virus was then washed away, and fresh culture medium and the exemplary fraction of this application were added, followed by culturing for 24 hours. The supernatant was collected, and the virus titer was determined by adding a dilution of the supernatant to susceptible cell lines.

[0421] Discussion: In mouse dermal fibroblasts, all three example fractions of this application significantly reduced HSV-1 titers at a concentration of 2000 μg / mL, and PBG-007-1 and PBG-002 showed similar effects at a concentration of 1000 μg / mL (see [link to study]). Figure 5 ).

[0422] b) Human skin fibroblasts (in vitro)

[0423] method:

[0424] Human skin fibroblast cell lines were pretreated for 24 hours with different concentrations of PBG-002, PBG-005, PBG-007-1, or a single culture medium. In the presence of the exemplary fraction of this application, virus was added at 0.1 MOI and the cells were cultured for 40 minutes. The virus was then washed away, and fresh culture medium and the exemplary fraction of this application were added, followed by culturing for 24 hours. The supernatant was collected, and the viral titer was determined by adding a dilution of the supernatant to susceptible cell lines.

[0425] Discussion: In human skin fibroblasts, PBG-005 significantly affected HSV-1 titers only at the highest concentration, while PBG-007-1 had a significant effect except at the lowest dose. PBG-002 significantly reduced HSV-1 titers at all concentrations (see [link to relevant documentation]). Figure 6 ).

[0426] Example 6: Effects on HSV-2

[0427] a) Mouse dermal cells (in vitro)

[0428] method:

[0429] Primary mouse dermal fibroblasts were pretreated for 24 hours with different concentrations of PBG-002, PBG-005, PBG-007-1, or a single culture medium. In the presence of the exemplary fraction of this application, virus was added at 0.1 MOI and the cells were cultured for 40 minutes. The virus was then washed away, and fresh culture medium and the exemplary fraction of this application were added, followed by culturing for 24 hours. The supernatant was collected, and the virus titer was determined by adding a dilution of the supernatant to susceptible cell lines.

[0430] Discussion: In the case of HSV-2 infection of mouse dermal fibroblasts, PBG-007-1 and PBG-002 had a significant effect on HSV-2 titers (see [link to discussion]). Figure 7 ).

[0431] b) Human skin fibroblasts (in vitro)

[0432] method:

[0433] Human skin fibroblast cell lines were pretreated for 24 hours with different concentrations of PBG-002, PBG-005, PBG-007-1, or a single culture medium. In the presence of the exemplary fraction of this application, virus was added at 0.1 MOI and the cells were cultured for 40 minutes. The virus was then washed away, and fresh culture medium and the exemplary fraction of this application were added, followed by culturing for 24 hours. The supernatant was collected, and the viral titer was determined by adding a dilution of the supernatant to susceptible cell lines.

[0434] Discussion: In human skin fibroblast cell lines infected with HSV-2, PBG-002 had no significant effect on HSV-2 titers. PBG-005 had a significant dose-dependent effect on HSV-2 titers, but this effect did not reach statistical significance. PBG-007-1 significantly reduced viral titers at the three highest doses, but the effect was not significantly dose-dependent (see [link to relevant documentation]). Figure 8 ).

[0435] c) Human foreskin fibroblasts (in vitro)

[0436] Primary human foreskin fibroblasts were pretreated for 24 hours with different concentrations of PBG-002, PBG-005, PBG-007-1, or a single culture medium. In the presence of the exemplary fraction of this application, virus was added at 0.1 MOI and the cells were cultured for 40 minutes. The virus was then washed away, and fresh culture medium and the exemplary fraction of this application were added, followed by culturing for 24 hours. The supernatant was collected, and the viral titer was determined by adding a dilution of the supernatant to susceptible cell lines.

[0437] Discussion: In the first study of HSV-2 titers in human foreskin fibroblast cultures, all three exemplary components of this application significantly reduced HSV-2 titers, but in different patterns. PBG-005 had a significant effect at a concentration of 2000 μg / mL, while PBG-002 significantly reduced viral titers at both concentrations. PBG-007-1 significantly reduced HSV-2 titers at a concentration of 1000 μg / mL, but not significantly at 2000 μg / mL. In the second study, both PBG-002 and PBG-007-1 caused a dose-dependent decrease in viral titers, particularly at 250 μg / mL and above. PBG-005 only affected HSV-2 titers at a concentration of 1000 μg / mL (see [link to study]). Figure 9A and 9B ).

[0438] d) Guinea pig model (in vivo)

[0439] method:

[0440] Female Hartley guinea pigs were randomly divided into 4 groups (N=5). The exemplary fraction of this application was resuspended in a stock solution of 5 mg / ml sterile water and incubated at 37°C for 1 hour. Animals were administered 10 mg of the compound daily via oral administration, orally dripped onto the tongue. Treatment was given 7 days before HSV-2 infection, on the day of infection, 12 days after infection (primary infection phase), and every other day after recovery from primary infection. 10 mg of the compound was also administered vaginally to the animals. 5 PFUHSV-2. Assess the animal's genital lesions, weight loss, urinary retention, hind limb paralysis, and overall condition daily. Use the following scale to score viral-induced genital skin lesions: 0, no obvious inflammation or infection; 1+, mild redness or swelling of the vaginal opening; 2+, one to four small blisters; 3+, five to nine large blisters; 4+, more than nine large ulcers with maceration; 5, hind limb paralysis. Additionally, guinea pigs are scored 0.5, 1.5, 2.5, or 3.5 when the lesion severity is moderate.

[0441] Animals were euthanized before the study was completed if they experienced a weight loss of more than 25%, a 4-point injury lasting more than 9 days, or complete hind limb paralysis or hind limb weakness that did not reverse within two days. All animals were euthanized on day 80 after HSV-2 challenge.

[0442] Ten days after HSV-2 IVAG challenge, vaginal secretions were collected from guinea pigs. The samples were cryopreserved, and viral titers were subsequently determined by plaque assay.

[0443] discuss

[0444] The viral titers in the PBG-005 and PBG-007-1 groups during primary infection were lower than those in the control and PBG-002 groups. The vaginal lesion score during primary infection was highest in the control group and lowest in the PBG-007-1 group.

[0445] The total cumulative injury score following primary infection was lowest in the PBG-002 and PBG-007-1 groups and highest in the control group. When plotting the cumulative injury number per individual animal, all animals in the control group (4 / 4; one animal in this group did not survive after primary infection) had vaginal lesions, compared to 3 / 5 animals in the PBG-005 group, 2 / 5 animals in the PBG-002 group, and 1 / 5 animals in the PBG-007-1 group (see [link to relevant documentation]). Figure 10 , 11 (as well as 12A and 12B).

[0446] Survival data are inconclusive because there were only 5 mice in each group, so each death resulted in a 20% decrease in survival. PBG-002 and PBG-007-1 had the same survival rate (80%), but PBG-007-1 maintained 100% survival for a longer period. The control group was the first to lose animals and had the fewest surviving animals at the end of the study (40%). PBG-005 had a moderate survival rate (60%) (see...). Figure 13 ).

[0447] Weight loss is a sign of serious illness in these animals, so their weight was recorded to determine if euthanasia was necessary. The PBG-007-1 group had the highest average weight (see [link]). Figure 14 ).

[0448] Although this application has been described with reference to examples, it should be understood that the scope of the claims should not be limited to the embodiments set forth in the examples, but should be given the broadest interpretation consistent with the entire description.

[0449] All publications, patents, and patent applications are incorporated herein by reference in their entirety, as if each individual publication, patent, or patent application were specifically and individually indicated to be incorporated herein by reference in its entirety. Where a term in this application is found to be defined differently in documents incorporated herein by reference, the definition provided herein shall serve as the definition of that term.

[0450] Full reference to the documents mentioned in the instruction manual

[0451] This document cites numerous publications. Full citations of these references are provided below. Each of these references is incorporated into this disclosure in its entirety by reference, as if each individual reference were specifically and individually indicated to be incorporated by reference.

[0452] Akira S, Uematsu S, Takeuchi O., Pathogen recognition and innate immunity, et al. Feb 24, 2006; 124(4): 783-801.

[0453] Hargreaves DC1, Medzhitov R., Innate sensory receptors for microbial infection, Journal of Clinical Immunology. November 2005; 25(6): 503-10.

[0454] Kawai T, Akira S, Antiviral signaling via pattern recognition receptors, Journal of Biochemistry. February 2007; 141(2): 137-45. Electronic version 2006

[0455] Philpott DJ1, Girardin SE., Roles of Toll-like receptors and Nod proteins in bacterial infection, Moore Immunology. Nov 2004; 41(11): 1099-108.

[0456] Seth RB, Sun L, Chen ZJ., Antiviral innate immune pathways, Cell Research. February 2006; 16(2):141-7.

[0457] Janeway CA Jr, Medzhitov R., Brief Introduction: The role of innate immunity in adaptive immune responses, serum immunity. October 1998; 10(5): 349-50.

[0458] Janeway CA Jr., The immune system evolved to be able to distinguish between infectious non-self and non-infectious self, modern immunity. January 1992; 13(1):11-6.

[0459] Janeway CA Jr, Medzhitov R., Damage to the host caused by lipoproteins, Current Biology. December 2, 1999; 9(23): R879-82.

[0460] Akira S, Takeda K., Toll-like receptor signaling, Nature Immunology (Revised). July 2004; 4(7):499-511.

[0461] O'Neill LA, How Toll-like receptors signal: What we know and what we don't know, current immune perspective. February 2006; 18(1):3-9.

[0462] Iwasaki A1, Medzhitov R., Toll-like receptor control of adaptive immune response, Natural Immunology. Oct 2004; 5(10): 987-95.

[0463] Bauer S1, Kirschning CJ H, Redecke V, Hausmann S, Akira S, Wagner H, Lipford GB., Human TLR9 confers responsiveness to bacterial DNA via species-specific CpG motif recognition. Proceedings of the National Academy of Sciences of the United States of America. July 31, 2001; 98(16):9237-42.

[0464] Krug A1, Towarowski A, Britsch S, Rothenfusser S, Hornung V, Bals R, Giese T, Engelmann H, Endres S, Krieg AM, Hartmann G., Toll-like receptor expression reveals CpG DNA as a unique microbial stimulant for plasmacytoid dendritic cells, synergistically inducing large amounts of IL-12 with CD40 ligand. European Journal of Immunology. October 2001; 31(10):3026-37.

[0465] Ashkar AA1, Rosenthal KL., Toll-like receptor 9, CpG DNA and innate immunity, Current Moore Medicine. Sep 2002; 2(6): 545-56.

[0466] Nguyen NH1, Nguyen CT, Pharmacological effects of ginseng on infectious diseases, Inflammatory Pharmacology. October 2019; 27(5):871-883.

Claims

1. A type of ginseng (American ginseng) Panax Quinquefolius A method for preparing acidic polysaccharide fractions from plant roots, the method comprising: At temperatures ranging from 10°C to 90°C, using C containing 50% to 95% by volume... 1-3 Extraction of American ginseng using a solvent mixture of alkyl OH and water ( Panax quinquefolius The root portion of the plant is used to produce a solvent mixture fraction and a first residue, wherein the ratio of the solvent mixture to the root portion is 20 to 1 volume weight to 1 volume weight. The first residue is fractionated from the solvent mixture; At a temperature of 10°C to 100°C, the first residue is extracted with water to produce an aqueous fraction containing an intermediate polysaccharide fraction and a second residue, wherein the ratio of water to the root portion is 20 to 1 volume weight to 1 volume weight. The aqueous fraction containing the intermediate polysaccharide fraction is separated from the second residue; The aqueous fraction is dried or concentrated to produce the intermediate polysaccharide fraction; The intermediate polysaccharide fraction is dissolved in water to produce a solution of the intermediate polysaccharide fraction, wherein the solution of the intermediate polysaccharide fraction is 1% to 10% by weight. C 1-3 Alkyl OH groups are added to the solution of the intermediate polysaccharide fraction to produce a neutral polysaccharide fraction and a first supernatant, wherein the C 1-3 The ratio of alkyl OH to the intermediate polysaccharide fraction solution is from 1:1 to 0.5:1 by volume. The neutral polysaccharide fraction is separated from the first supernatant to produce the neutral polysaccharide fraction; C 1-3 Alkyl OH is added to the first supernatant to produce a second supernatant rich in acidic polysaccharide fractions, wherein C 1-3 The ratio of alkyl OH to the first supernatant is from 0.5:1 to 8:1 by volume; The acid-rich polysaccharide fraction was separated from the second supernatant. The acid-rich polysaccharide fraction is dissolved in water to produce a solution rich in acid-rich polysaccharide fraction, wherein the solution rich in acid-rich polysaccharide fraction is 1% to 10% by weight. C 1-3 Alkyl OH groups are added to the solution rich in the acidic polysaccharide fraction to produce a precipitate and a third supernatant containing the acidic polysaccharide fraction, wherein C 1-3 The ratio of alkyl OH to the solution rich in acidic polysaccharide fraction is from 1:1 to 0.5:1 by volume. The third supernatant containing the acidic polysaccharide fraction is separated from the precipitate. The acidic polysaccharide fraction is used to prepare drugs for treating influenza A virus infection or herpes simplex virus infection.

2. The method according to claim 1, further comprising: The third supernatant is dried or concentrated to produce the acidic polysaccharide fraction; or The solution rich in acidic polysaccharide fractions was loaded onto a chromatographic column containing anion exchange resin; and The acidic polysaccharide-rich elution fraction is produced by eluting the anion exchange resin with an ammonium acetate buffer to produce the acidic polysaccharide elution fraction. The elution includes a first step in which the buffer is 10 mM to 100 mM ammonium acetate, and a second step in which the buffer contains 0.5 M to 1.5 M ammonium acetate.

3. The method according to claim 2, further comprising: The acidic polysaccharide elution fraction is dried or concentrated to produce the acidic polysaccharide fraction.

4. The method of claim 1, wherein the solvent mixture in the step of extracting the root portion comprises 80% by volume C 1-3 Alkyl OH to 90% C by volume 1-3 Alkyl OH.

5. The method of claim 1, wherein the ratio of the solvent mixture to the root portion is 10 to 1 by volume weight.

6. The method of claim 1, wherein in the step of extracting the first residue, the ratio of water to the root portion is 12 to 1 by volume weight.

7. The method according to claim 1, wherein the American ginseng is extracted using the solvent mixture. Panax quinquefolius The step of extracting the root portion of the plant or the step of extracting the first residue with water is carried out at the following temperatures: 20°C to 100°C, 30°C to 100°C, 40°C to 100°C, 50°C to 100°C, 60°C to 100°C, 70°C to 100°C, 80°C to 100°C or 90°C to 100°C, and for more than 1 hour, more than 2 hours or more than 3 hours.

8. The method according to any one of claims 1 to 7, wherein C is... 1-3 In the step of adding alkyl OH to the solution of the intermediate polysaccharide fraction, C 1-3 The ratio of alkyl OH to the solution of the intermediate polysaccharide fraction is from 1:1 to 0.5:1 by volume.

9. The method according to any one of claims 1 to 7, wherein C 1-3 Alkyl OH in C 0.5:1 to 6:1 volume ratio 1-3 The ratio of alkyl OH to the first supernatant is added to the first supernatant.

10. The method according to any one of claims 1 to 7, wherein the solution rich in acidic polysaccharide fraction or the solution of intermediate polysaccharide fraction is 5% by weight.

11. The method according to any one of claims 1 to 7, wherein C 1-3 Alkyl OH in C at a volume ratio of 0.7:1 to 0.5:1 1-3 The alkyl OH group is added to the solution rich in acidic polysaccharide fraction at a ratio of alkyl OH to the solution rich in acidic polysaccharide fraction.

12. The method according to any one of claims 1 to 7, wherein each C 1-3 The alkyl OH group represents ethanol.

13. The method according to any one of claims 1 to 7, wherein the buffer in the first step of elution is 20 mM ammonium acetate, and the buffer in the second step of elution is 1 M ammonium acetate.

14. The method according to any one of claims 1 to 7, wherein the acidic polysaccharide fraction is used to prepare a medicament for treating influenza A virus infection.

15. The method of claim 14, wherein the influenza A virus infection is an H1N1 virus infection.

16. The method according to any one of claims 1 to 7, wherein the acidic polysaccharide fraction is used to prepare a medicament for treating herpes simplex virus infection.

17. The method of claim 16, wherein the herpes simplex virus infection is herpes simplex virus 1 (HSV-1) infection or herpes simplex virus 2 (HSV-2) infection.

18. An acidic polysaccharide fraction produced by the method of any one of claims 1 to 13, for use in the preparation of a medicament for treating influenza A virus infection or herpes simplex virus infection.

19. The acidic polysaccharide fraction according to claim 18, wherein the acidic polysaccharide fraction is used to prepare a medicament for treating influenza A virus infection.

20. The acidic polysaccharide fraction according to claim 19, wherein the influenza A virus infection is an H1N1 virus infection.

21. The acidic polysaccharide fraction according to claim 18, wherein the acidic polysaccharide fraction is used to prepare a medicament for treating herpes simplex virus infection.

22. The acidic polysaccharide fraction according to claim 21, wherein the herpes simplex virus infection is herpes simplex virus 1 (HSV-1) infection or herpes simplex virus 2 (HSV-2) infection.

23. An acidic polysaccharide fraction comprising 2 wt% to 5 wt% terminally linked rhamnopyranosyl residues (t-Rha), 3 wt% to 6 wt% terminally linked arabinose-furanosyl residues (t-Araf), 1 wt% to 4 wt% 2-linked rhamnopyranosyl residues (2-Rha), 3 wt% to 6 wt% terminally linked glucopyranosyl residues (t-Glcp), 12 wt% to 15 wt% terminally linked galactopyranosyl residues and terminally linked galacturonic acid-pyranosyl residues (t-Galp and t-Gal A) 4% to 8% by weight of 4-linked arabinopyranosyl residues or 5-linked arabinofuranosyl residues (4-Arap or 5-Araf), 1% to 4% by weight of 2,4-linked rhamnopyranosyl residues (2,4-Rha), 1% to 4% by weight of 3-linked galactopyranosyl residues (3-Galp), 31% to 35% by weight of 4-linked galactopyranosyl residues and 4-linked galacturonic acid pyranosyl residues (4-Galp and 4-GalA), 22% to 26% by weight of 4-linked glucopyranosyl residues (4-Glcp), 0.5% to 3% by weight of 2,4-linked galactopyranosyl residues (2,4-Galp), and 0.5% to 3% by weight of 3,6-linked galactopyranosyl residues (3,6-Galp). The acidic polysaccharide fraction is used to prepare drugs for treating influenza A virus infection or herpes simplex virus infection.

24. The acidic polysaccharide fraction according to claim 23, wherein the acidic polysaccharide fraction is used to prepare a medicament for treating influenza A virus infection.

25. The acidic polysaccharide fraction according to claim 24, wherein the influenza A virus infection is an H1N1 virus infection.

26. The acidic polysaccharide fraction according to claim 23, wherein the acidic polysaccharide fraction is used to prepare a medicament for treating herpes simplex virus infection.

27. The acidic polysaccharide fraction according to claim 26, wherein the herpes simplex virus infection is herpes simplex virus 1 (HSV-1) infection or herpes simplex virus 2 (HSV-2) infection.

28. A composition comprising the acidic polysaccharide fraction of claim 18 or 23 and a carrier, said composition for preparing a medicament for treating influenza A virus infection or herpes simplex virus infection.

29. The composition of claim 28, wherein the composition is used to prepare a medicament for treating influenza A virus infection.

30. The composition of claim 29, wherein the influenza A virus infection is an H1N1 virus infection.

31. The composition of claim 28, wherein the composition is used to prepare a medicament for treating herpes simplex virus infection.

32. The composition of claim 31, wherein the herpes simplex virus infection is herpes simplex virus 1 (HSV-1) infection or herpes simplex virus 2 (HSV-2) infection.

33. The composition according to any one of claims 28 to 32, wherein the composition is a pharmaceutical or nutritional product composition comprising a pharmaceutically acceptable carrier.

34. Use of the acidic polysaccharide fraction of claim 18 or 23 in the preparation of a medicament for treating influenza A virus infection or herpes simplex virus infection.

35. The use according to claim 34, wherein the acidic polysaccharide fraction is used to prepare a medicament for treating influenza A virus infection.

36. The use according to claim 35, wherein the influenza A virus infection is an H1N1 virus infection.

37. The use according to claim 34, wherein the acidic polysaccharide fraction is used to prepare a medicament for treating herpes simplex virus infection.

38. The use according to claim 37, wherein the herpes simplex virus infection is herpes simplex virus 1 (HSV-1) infection or herpes simplex virus 2 (HSV-2) infection.

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