JP2000072680A - Immunological function controller - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain an immunological function controller that has a low molecular weight with reduced side-effects and is useful for diseases, for example, malignant tumor, rheumatoid arthritis, atopic dermatitis, diabetes and the like by subjecting L(+)-lactic acid to dehydration condensation in a nitrogen gas atmosphere by stepwise decompression and stepwise temperature rise. SOLUTION: L(+)-lactic acid is subjected to dehydration condensation reaction through stepwise decompression and stepwise temperature rise in a nitrogen gas atmosphere to give cyclic type and/or chain type oligomers represented by the formulas (C3H4O2)z and [(C3H4O2)z-H2O] (z=2-23) and having two types of molecular structures given by formula I and formula II of a zigzag type cyclic one and of a clathrated almost zigzag type C-letter shaped chain one. The ratio of the cyclic type to the chain one is about 7:3. The daily dose is about 4 g at minimum.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

[0001] The present invention relates to an immune function modulator for use in intractable diseases accompanied by abnormal immune functions, such as malignant tumors, rheumatoid arthritis, atopic dermatitis and diabetes in animals including humans.

[0002]

2. Description of the Related Art Conventionally, as typical substances or means for restoring or enhancing a biological defense function or an immune response and thereby obtaining a therapeutic effect, cytokines such as interferon and interleukin, vaccines, in v.
There are activation of immunocompetent cells such as adoptive immunotherapy in which lymphocytes have been activated in vitro, and immunoadjuvants or immunomodulators such as polysaccharides and bacterial components.

[0003] However, for intractable diseases accompanied by abnormal immune functions, most of them are in the research stage, and few of them have been confirmed to be clinically useful. Furthermore,
Since many of these compounds have high molecular weight and complex biological activities, it is difficult to exert a certain effect on immune function, and it is effective, ineffective, or uncertain, It has been pointed out that it is difficult to control the amount, that there is a risk of serious and various side effects, that antibodies appear after use, that the decomposition is fast, and the like.

At present, no low molecular weight substance which does not have such disadvantages and which regulates the immune dysfunction observed in intractable diseases by oral ingestion has not been found.

On the other hand, L-lactic acid is heated at normal pressure or reduced pressure in an atmosphere of an inert gas such as nitrogen gas, and the resulting reaction solution is dissolved in methanol or ethanol while heating, and then filtered. After drying, dissolve in acetonitrile or directly dissolve the solution in acetonitrile at 25% of pH 2-3 beforehand.
Reversed phase ODS equilibrated with acetonitrile aqueous solution
Alternatively, perform chromatography on a DS column,
Elute with 30-50% acetonitrile aqueous solution
2 to 3 fractions eluted with an aqueous solution having an acetonitrile concentration of 70% or more, wherein the L-lactic acid linear condensate having a condensation degree of 5 to 23 and the L-lactic acid cyclic condensate having a condensation degree of 2 to 15 An agent for suppressing the growth of malignant tumor cells in animals, including humans, comprising a mixture of the above compounds has been proposed in JP-A-5-310581.

[0006]

According to the present invention, a cyclic oligomer molecule (C ₃ H ₄ O ₂ ) _z and a chain type oligomer molecule ((C ₃ H ₄ O ₂ ) _z -H ₂ O) are about 7 The objective of the present invention is to provide an L (+)-lactic acid oligomer having an immune function regulating action contained in a ratio of 3:

[0007] This substance is administered orally to patients suffering from intractable diseases accompanied by immune dysfunction such as malignant tumors, rheumatoid arthritis, atopic dermatitis, diabetes, etc., to give liver, digestive and hematopoietic functions. To activate the body's immune system and to alleviate symptoms specific to each disease. These effects act on the function of antigen presenting cells such as macrophages by affecting the molecular expression of the receptor or the membrane structure, thereby activating the ability to induce T cell differentiation, and producing cytokines such as IL-2. This was considered to be the result of normal maintenance / regulation of the functions of a series of immunocompetent cells including NK cells and multinuclear neutrophils.

[0008] Such an immunomodulatory effect by oral ingestion is a new finding that has not been known so far, and rheumatoid arthritis, periarteritis nodosa, scleroderma, ulcers, to which autoimmunity is not least involved. Intractable diseases such as ulcerative colitis and Crohn's disease, malignant tumors whose immune function plays an important role in the occurrence, progression and prognosis of the disease, various intractable diseases such as atopic dermatitis, diabetes, and AIDS It is expected to work effectively and eventually lead to healing and contribute to the improvement of modern medical care.

[0009]

In order to achieve the above object, the present invention provides a cyclic and chain type oligomer obtained by dehydrating and condensing L (+)-lactic acid in a nitrogen gas atmosphere by stepwise decompression and heating. Substances whose chemical compositions are (C ₃ H
₄ O _{2) z,} and _{{(C 3 H 2 O)} z -H 2 O} ( here
_z = 2 to 23), and uses an immune function modulator comprising two kinds of molecular structures, a zigzag cyclic structure and a clathrate-like substantially zigzag C character chain structure.

[0010]

Embedded image

Chemical composition of the synthesized product From the results of FD-MS analysis, the synthesized L (+)-lactic acid oligomer contains cyclic type and chain type in a ratio of about 7: 3, and the degree of polymerization z is up to 23. Is a condensation polymer. From this, the present inventors have proposed that this substance be converted to cyclic polylactic acid (Cyclic).
poly-lactate: hereinafter abbreviated as CPL)
It was named.

[Table 1]

Three-dimensional structure model based on energy calculation CPL molecules show a hollow zigzag closed ring (FIG. 1) at a low degree of polymerization based on the energy calculation of molecular dynamics using a computer model. The degree of polymerization shows a zigzag structure in which the closed oblong ring is bent like a C-shaped character (FIG. 2), and has both annular and chain characteristics. Chain type oligomer molecule {(C ₃ H ₄ O)
₂₎ z _{-H 2 O}} shows the structure for connecting the two-dimensional direction or solidified or elongation linked in a chain.

When the CPL obtained by the present invention is actually used as an oral preparation, it can be used as it is. However, in consideration of the properties of the substance, usually, a stabilizer such as calcium lactate, calcium carbonate, Mannitol,
Sorbitol and the like are added. It can be administered in the form of a mixture with other pharmacologically active substances, and the dosage form may be suitably selected from powders, granules, tablets, dragees, capsules, suspensions, emulsions and the like. The dose of CPL varies depending on the method of administration, type of disease, disease state, stage, etc., but the minimum dose is about 4 g per day, and it may be necessary to administer this dose several times over.

Formulation Example 1800 g of CPL of the present invention is added to 500 g of sorbitol
And 200 g of calcium carbonate, solidify and pulverize to a fine powder, and aseptically fill the container.

[0015]

BEST MODE FOR CARRYING OUT THE INVENTION Production Example
Add (+)-lactic acid 2500ml and nitrogen gas 500ml
/ Min., And while the distillate was being introduced into a flask equipped with a reflux condenser, the mixture was heated to 145 ° C for 3 hours to distill off free water, and then reduced to 150 mmHg and heated for 3 hours. The mixture was heated at 5 mmHg for 3 hours and heated to 170 ° C., and finally heated to 190 ° C. for 1.5 hours to obtain CPL as a dehydrated condensate.

Effects on the immune function of patients suffering from intractable diseases NK (natural killer) cells mainly exist in blood,
It acts on target cells spontaneously without antigen sensitization, has antitumor properties and regulates antibody production, and is considered to be involved in maturation and differentiation of bone marrow cells. The NK cell activity and ADCC (antibody-dependent cell mediated injury) activity possessed by NK cells are useful for judging immune function status. Therefore, subjects suffering from multiple intractable diseases such as anemia, liver cirrhosis, rheumatism, and Hashimoto's were given 10 g of CPL once daily in the early morning for 2 weeks, and before and 1 and 2 weeks after taking. NK cell activity and ADCC activity were measured, and a general blood test including immunoglobulin was performed. As a result, CP
L administration clearly improved anemia, which was ineffective with existing therapeutic agents, and increased blood albumin (11
A / G ratio and total protein content were increased (140% and 104%, respectively). In terms of immunobiochemistry, in addition to the increase in IgE (305%), NK cell activity was maintained at a high value, and ADCC activity was slightly increased (109%).
%)showed that. Therefore, it was considered that CPL acts on the liver function, hematopoietic function, etc., and activates the biological immune system or the biological defense system.

Clinical treatment results by oral administration Case 1: Multiple rheumatoid arthritis (female, 53 years old) From around 1975, she was diagnosed with multiple rheumatoid arthritis and was treated. Deformation of fingers and footprints has occurred since 1990 or 3rd. He received treatment from that time but did not turn around and had moderate pain. C in combination with Zyrolic from December 1997
PL 10 g / day was taken once a day in the early morning. At present, although there is moderate deformation, remarkable reduction of pain and disappearance of pain during movement are observed, and the course is good.

Case 2: Knee arthritis (male, 67 years old) Since about 1988, there was occasional pain when moving, and orthoses such as supporters were used. Pain intensifies especially in cold weather, taking voltaren during pain and using herpes as an external medicine. Take CPL 10g / day from April 1996. Pain and swelling disappeared from around February 1997, and CPL
You are taking.

Case 3: Rheumatoid arthritis (female, 59 years old) Hospitalized in November 1994, taking gyrolic. Heisei 9
She has been taking CPL 6 g / day since November 2011, and now she has stopped using gyrolic because her pain has disappeared, and her progress has been good.

[0020]

According to the results of mass spectrometry and structural analysis of the CPL of the present invention, it has been found that the CPL has a cyclic and chain type molecular structure and is an oligomer substance which easily reacts with other molecules. It was confirmed that it plays an important role in improving and regulating functions. So far, the antitumor effect of this substance, the inhibitory activity of anaerobic glycolysis, the improvement of digestive function and the effect on glycolipid metabolism have been clarified. The structural properties and biological activity of these CPLs can be said to be the properties first found in carbohydrate oligomers of (CH ₂ O) _n composition, and they are also biocompatible and biocompatible without impurities synthesized without using a catalyst. Considering that it is a degradable oligomer and has almost no side effects after administration, it is suggested that this substance is similar to a physiologically active substance in the living body, or a substance indispensable for maintaining biological function and homeostasis. Suggests. In particular, immunity is one of the key and complex functions in maintaining life, and many intractable diseases for which effective treatment is not available are accompanied by considerable immune function abnormalities, and thus, the present invention provides immunity. It is expected that the normal maintenance / regulatory action of CPL on the immune function can be widely applied to these intractable diseases and lead to its improvement. Furthermore, the mechanism of action of CPL has features not found in existing pharmaceuticals. That is, CPL is present in M (multifol) present in the intestinal Peyer's patch after ingestion.
d) After being taken up by cells, it modulates the response of T cells through the action on antigen presenting cells such as macrophages, and works in conjunction with the control of cell mediators such as cytokines and the functions of the endocrine and nervous systems. It has been shown to activate systemic cellular and humoral immune responses, and it is thought that oral administration of CPL exerts an immune function-modulating effect by at least such a mechanism. It is also known that the intestinal flora and its producing substances, short-chain fatty acids such as propionic acid, butyric acid, and lactic acid, are involved in the biological defense function, and L (+)-lactic acid is used as a starting material for these functions. C
It is fully conceivable that PL is acting effectively. C
Although PL alone can provide a sufficient effect, it is possible to expand the application range and improve the therapeutic effect by imparting other pharmacological actions or using it in combination with existing drugs. As described above, the CPL of the present invention is a low molecular weight oligomer substance having useful biological activities such as acting on the immune function of a living body without side effects, and having a cyclic structure as a main molecular structure. This can be said to bring new developments in the treatment and prevention of various diseases mainly including intractable diseases.

[Brief description of the drawings]

FIG. 1. Molecular structure model of L (10) -lactic acid oligomer at low polymerization degree

FIG. 2: Molecular structure model of L (+)-lactic acid oligomer at high polymerization degree

Claims (1)

[Claims] 1. A cyclic and chain type oligomer substance obtained by dehydrating and condensing L (+)-lactic acid in a nitrogen gas atmosphere by stepwise decompression and temperature elevation, each having a chemical composition of (C ₃ H)
₄ O ₂₎ are _z and _{{(C 3 H 2 O)} z -H 2 O} ( where z = 2 to 23), substantially molecular structure of zigzag ring structure and clathrate-like zigzag-C character shape chain An immune function modulator comprising two types of structures.