JPH0317010A - Preventive for dental caries - Google Patents

Abstract

PURPOSE:To obtain a preventive for dental caries having proliferation-inhibiting effect against Streptococcus mutans by extracting KANJU (rhizome of Dryopteris crassirhizoma) with methanol, fractionating the extract with a dialysis tube and using the obtained fraction having a specific molecular weight as an active component. CONSTITUTION:KANJU is extracted with methanol and the components soluble in a non-hydrophilic organic solvent are removed from the extract to obtain an aqueous solution. The solution is put into a dialysis tube having a fractional molecular weight of 1,000 and dialyzed with water. The inner liquid is put into a dialysis tube having a fractional molecular weight of 10,000 and dialyzed with water to fractionate into an inner liquid and an outer liquid and obtain NPF-87KAN-I having a molecular weight of 1,000-10,000 and NPF-87KAN-II having a molecular weight of >=10,000. Both fractions are separately subjected to high-performance liquid chromatography and each fraction is separated into two fractions (IA having a molecular weight of 8,100, IB having a molecular weight of 4,900, IIA having a molecular weight of 12,800 and IIB having a molecular weight of 9,400). KAN-I is a mixture of IA and IB at a ratio of about 1:6 and KAN-II is a mixture of IIA and IIB at a ratio of about 1:4. Both fractions have comparable activities for the inhibition of the growth of Streptococcus mutans.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Field of Application] The present invention relates to a caries preventive agent having an effect of inhibiting the growth of dental caries bacteria.

[Conventional technology]

Since the chemical bacterial tax proposed by W. D. Miller at the end of the 19th century, various debates have been held regarding the cause of dental caries, but currently the Lactobacillus theory has been the most widely believed for many years. was rejected and replaced by Streptococcus mutans (Streptococcus mutans).
ccus mutans; hereinafter referred to as S. It's called mutans. ) is thought to be the main causative bacteria.

On the other hand, conventional caries prevention methods include: l) Completely removing sucrose from food, 2) Strengthening tooth structure, and 3) Preventing the production of sticky glucan, which causes plaque formation. 4) Measures to inhibit the growth of dental caries bacteria were known.

[Problem that the invention seeks to solve]

However, all of the above caries prevention methods were incomplete. For example, it is unrealistic to remove sucrose from food, and dentin strengthening agents, adhesive glucan inhibitor enzymes, bactericides, etc. have drawbacks such as side effects or low effectiveness. It wasn't something to be satisfied with.

Therefore, there has been a desire for a caries preventive agent that has no side effects, can be produced at low cost, and is highly effective.

[Means to solve the problem]

Under these circumstances, the present inventors conducted intensive research in search of a natural product that is highly safe and can be produced at low cost. The present inventors have discovered that an oral cavity composition can be obtained which is recognized to be effective and can prevent dental caries, and the present invention has been completely abandoned.

That is, the present invention provides an effective anti-caries agent using the methanol extract of Oriental caries.

The present invention has a molecular weight of 1,000 obtained by fractionating a defatted methanol extract of the sucrose using a dialysis tube.
The object of the present invention is to provide a caries preventive agent which is effective in using a methanol extract of 10,000 to 10,000 fractions.

In addition, the present invention provides a methanol extract of defatted solids with a molecular weight of 10,000 obtained by fractionation from a dialysis tube.
The object of the present invention is to provide a caries preventive agent whose effective ingredient is a methanol extract of the above-mentioned 100% citrate. The present invention also provides NPF8 7KAN I, NPF8 7KAN I, and NPF 8KAN I, which have the following physicochemical properties and are obtained by further purifying the above extract.
7KAN II, NPF-8 7KAN-IA
SNPF -8 7KAN IB, NPF -
8 7KAN-I and NPF -8 7KAN-I
The present invention provides a caries preventive agent containing a substance selected from the group consisting of IB as an active ingredient.

(NPF-87KAN-1) (i>Shape: Light brown powder (ii) Melting point: No clear melting point or decomposition point.

(iii) Elemental analysis (iv) Molecular weight: l, 000 to 10,000 (by dialysis tube) (v) Infrared absorption spectrum: 1443, 1375, 1284, l245, l103,
l067, 779 (vi) Ultraviolet absorption spectrum; 305 shoulder (107 7) 510 shoulder (21.6) (vii) Solubility: Soluble in water, methanol, ethanol.

Insoluble in hexane, ether, ethyl acetate, and chloroform.

(vin) Color reaction: (v) Infrared absorption spectrum; (vi) Ultraviolet absorption spectrum: (NPF 8 7KAN If) (i) Shape: Light brown powder (ii) Melting point: No clear melting point or decomposition point.

<iii) Elemental analysis: 510 shoulders (25.5) (vj) Solubility: Soluble in water, methanol and ethanol.

Insoluble in hexane, ether, ethyl acetate, and chloroform.

(vji) Color reaction: (iv > molecular weight: 10,000 or more (by dialysis tube) (blade PF-87KAN-IA) (i
) Shape: Light brown powder (ii) Melting point: No clear melting point or decomposition point.

(iii) Elemental analysis: C: 37.71% H: 3.97% N: 2.83% Ash: 16.48% (iv) Molecular weight: 8,100 (Gel permeation 1x chromatography based on polyethylene glycol) (v) Infrared absorption spectrum: 1445. 1384. 1259. 1114,10
67. 802 (vi) Ultraviolet absorption spectrum: (vii) Solubility: Soluble in water, methanol, and ethanol.

Insoluble in hexane, ether, ethyl acetate, and chloroform.

(vii) Color reaction: Ferric chloride positive ninhydrin negative p-anisidine-phthalate negative anisidine-diphenylamine negative Dragendorff negative (NPF-87KAN-IB) (i) Shape:
Light brown powder (ii) Melting point: No clear melting point or decomposition point.

(iii) Elemental analysis: C: 45.78% H: 4.45% N: 0.71% Ash: 10.45% <0.9% (iv> Molecular weight: 4,900 (based on polyethylene glycol) (v) Infrared absorption spectrum: 1442.1284, 1262.1206.11
57, 1113. 1067. 803(v1) Ultraviolet absorption spectrum: (vj) Solubility: Soluble in water, methanol, and ethanol. Insoluble in hexane, ether, acetic acid, ethinobe, and chloroform.

(viii) Color reaction: Ferric chloride Positive Ninhydrin Negative p-anisidine-phthalate Negative Anisidine-Diphenylamine Negative Dragendorff Negative (NPF-8 7KAN-I [A) (i) Shape:
Light brown powder (ii>M point; no clear melting point or decomposition point.

(iii) Elemental analysis: C:・3 8. 6 6% H: 3.67% N: 2.83% Ash: 22.94% (iv) Molecular weight: 12,800 (Based on gel permeation chromatography using polyethylene glycol as a standard.) (v) Infrared absorption spectrum : 1372, 1283. 1205, 1113.79
5 (vi) Ultraviolet absorption spectrum: Water λ. ．． ．． nm (E+c+a) 280(
vj) Solubility: Soluble in water, methanol and ethanol. Insoluble in hexane, ether, ethyl acetate, and chloroform.

(vii) Color reaction: Ferric chloride Positive ninhydrin Negative p-anisidine-phthalic acid Negative anisidine-diphenylamine Negative Dragendorff Negative (NPF-8 7KAN n B) (i>Shape:
Light brown powder (ii) Melting point: No clear melting point or decomposition point.

(iii) Elemental analysis: C: 44.97% H: 4, 44% N: 0.79% Ash: 14.32% (iv) Molecular weight +9. 400 (Based on gel permeation chromatography using polyethylene glycol as a standard) (v) Infrared absorption spectrum: 1445, 1379. 12B5. 1261.12
02. 1096. 802 (vi) Ultraviolet absorption spectrum: (vj) Soluble Soluble in water, methanol and ethanol.

Insoluble in hexane, ether, ethyl acetate, and chloroform.

(viii) color reaction; Ferric chloride Positive Ninhydrin Negative p-anisidine-phthalate negative Anisidine-diphenylamine negative Dragendorff Negative The present invention will be explained in more detail below.

■． Raw materials As a raw material for the caries preventive agent of the present invention, caries can be used, but it is preferable to use those that have been subjected to treatments such as drying, coarse crushing, and pulverization so that they can be easily processed and extracted.

It is also convenient to use commercially available crude drugs.

The following is also known about Kanshu.

In other words, the decoction of Kanshu has a remarkable insecticidal effect on insects such as worms, roundworms, earthworms, and leeches in vitro. In addition, this decoction has a strong intestinal and uterine contraction effect, has an initial stimulating effect on the respiratory center, and after a while has the effect of suppressing cardiac activity and lowering blood pressure. It is also known to have antifungal effects on the skin and, to a lesser extent, to constrict blood vessels.

Furthermore, Kanshu decoction is known in the veterinary field to be effective in treating liver fluke disease and other parasitic diseases in cattle.
Even if the decoction is diluted to 1/3,000, it has a strong contractile effect on the excised uterus of domestic rabbits. Kanshu (also called kanzhong) is a herbal medicine made from various protozoa, and the following protoplants and production areas are currently recognized in market products in Japan, China, and the Korean Peninsula.

(Basic source) 1) Oshida science (Apicliaceae)
) of Dryopteris crassirhi
zoma NAKAI. 2>Osidae (^spid)
iaceae) thyrium pycnosrum CHRIST.
(=A.acrost+cho+des Dr+
ats. =Lunathyrium Pycnos
orum Ko+nz. )． Ezomesida (Mesida, Kouraimesida) A, brevifrons L
KAr exKITAGAWA+ 3) B lechnaceae Bl
eehum orientate L. ．． 4) Woodv+ardia unigemmata N of Blechnaceae
AKAI. :] Mochishida W. orientalis
3w,, 5) Chirindae (mouth smundaceae)
ae＞ mainspring Osmunda japonica
THU)18. , 6) Aspididae (Aspidiaceae)
aceae)'s cycad! Jetteucciast
ruthiopteris To. ．． Ro (made in China). 7) Struthiopte of the family Blechnaceae
ris nipponica (Ku, lzx) LKA
+ (made in Japan), dried rhizomes of the above plants.

(Produced!!) 1) It is produced in the Northeastern Islands of China and the Korean Peninsula, and is sold in the Tohoku, Beijing, Tianjin, Shenyang (formerly Mukden), Lunda (formerly Dalian), and Seoul (formerly Gingcheng) markets.

2) Produced in North China, Central China, etc., and consumed in those regions, it also appears in the Xuanbei and Beijing markets. 3) Produced in South China and sold in the Guangdong market. 4) Found throughout East China, South China, and Sichuan provinces. 5) It is produced in East China, Central China, and Henan and is consumed in these areas. This product is sold in the Shanghai market. 6) Produced in Hebei and other provinces of northern China, and sold in markets such as Beijing. 7) Produced all over Japan.

■． Extraction of active ingredients a) NPF-8 7KAN-I and NPF
-87KAN-II NPF-87KAN-I and NPF- which are active ingredients of the caries preventive agent of the present invention
8 7KAN-II is a phenolic substance and is characterized by its growth-inhibiting activity against dental caries. Therefore, it can be purified appropriately using this growth-inhibiting activity as an indicator by extraction with water, methanol, methanol such as ethanol, centrifugation, filtration, etc. It can be isolated and purified by applying various methods.

These methods can be applied singly, in combination in any order, or repeatedly as needed. Below NPF -
8 7KA? i-I and NPF-87KAN
An example of a method for extracting -H will be explained.

In addition, the raw material Kanshu is dried and dried to make it easier to process and extract.
It is preferable to use a material that has been subjected to processing such as coarse crushing or pulverization.

b) Using a degreasing solvent such as hexane or ether, the hexane- or ether-soluble portion of the raw material is extracted and removed at room temperature or by heating.

(1) The raw material from which the portion soluble in a degreasing solvent such as hexane or ether has been extracted and removed is air-dried or vacuum-dried to remove the solvent.

C> Next, methanol is added to the extraction raw material and extracted according to a conventional method. Usually extracted under boiling, but at room temperature, 4℃
Even if extraction is carried out in a cold room, active ingredients can be obtained.

2) After concentrating the obtained methanol extract to dryness, extract it with water. Specifically, water is added and suspended, insoluble matter is filtered out, water is further added to the insoluble matter, stirred well, filtered, and combined with the previous solution.

e) Add an equal amount of a non-hydrophilic organic solvent such as ethyl acetate or chloroform to this aqueous solution and remove the organic solvent soluble portion.

f) The aqueous layer from which the non-hydrophilic organic solvent soluble portion has been removed is placed in a dialysis tube (Spectra/Pore 6; manufactured by Spectrum Medical Industries) with a molecular weight cut off of 1,000, and dialyzed against water to remove the internal solution. and the external fluid.

g) The dialysis fluid fractionated with a dialysis tube with a molecular weight cutoff of 1,000 was further placed in a dialysis tube with a molecular weight cutoff of 10.000 (Svectra/Pore 6; manufactured by Spectrum Medical Industries) and submerged in water. Dialyze and separate into internal and external fluids.

h) Molecular weight fractionated in this way1. 0 0 0~t
Fractions of o. Each of these fractions can be obtained as a light brown powder by further operations such as freeze-drying.

The present inventor has determined that active substances fractionated with molecular weights of 1,000 to 10,000 and 10,000 or more are used as NPF, respectively.
-87KAN-I and NPF-87K
It was named AN-It.

b) NPF- 8 7KAN- I ASNPF-
8 7KAN- IB,NPF- 8 7KAN-
n A and NPF-8 7 KAN-II B In this way, NPF-8 was fractionated using a dialysis tube.
7KAN I and NPF - 8 7KAN -
II can be further separated into two fractions each.

That is, as described later, NPF 87KAN I is a fraction with a molecular weight of 8,100 (NPF-87KAN
-IA) and the fraction with a molecular weight of 4,900 (NPF-8
7KAN-IB), also NPF-8 7KAN
- I[ is the fraction with a molecular weight of 12.800 (NPF -
87KAN-I[A] and a fraction with a molecular weight of 9,400 (NPF-87KAN-IIB) (see Figures 11A and B).

These four types (NPF-8 7KAN-IA,
NPF-87KAN =IBS NPF-87K
AN-mA, and NPF-8 7KAN-
The separation and purification of the substance IIB) can be carried out by various known methods, but it is preferably carried out using a high performance liquid chromatograph under the following conditions.

(i) Separation force ram In this case, a separation force ram filled with a distribution/adsorption type resin, an ion exchange resin, a gel filtration type separation agent, etc. can be used. It is also preferable to additionally use a device for automatic injection and automatic fractionation.

(ii) Eluent As the eluent, in addition to the water-methanol system, various buffer solutions such as water, acetonitrile, dimethylformamide, acetic acid, butanobehexane, etc. can be used alone or mixed in any ratio. can.

(iii) Indicator As an indicator for detecting the effective substance of the present invention, 28
Absorbance at a wavelength of 0 nm and activity to inhibit growth of dental caries bacteria can be used.

(iv) Processing method As the processing method, an oven column, medium pressure or high pressure method can be used.

NPF 8 7KAN I A, NPF
8 7KAN-IB, NPF -8 7KAN -
IIA and NPF-87KAN-I[B each showed a single peak in the high-performance liquid chromatography, and their activities to inhibit the proliferation of dental caries bacteria matched.

In addition, gel permeation chromatography performed to measure the molecular weight of this substance shows an i-peak.

When examined using the standard molecular weights of various polyethylene glycols, the molecular weight of NPF-87KAN-IA was 8,100, while that of NPF-87KAN-IB was 4.
90 0, NPF-8 7KAN-IrA is 1 2,
80 0 and NPF -8 7KAN -JIB is 9
．． It was decided to be 4QO.

As mentioned above, NPF - 8 7 KAN -
I is NPF-8 7KAN-I A and NPF
- a mixture of 8 7KAN-IB, while NPF 8
7KAN If is NPF-8 7KAN-I
It is a mixture of IA and NPF-87KAN-I[B. Analysis of high-performance liquid chromatograms revealed that NPF
-8 7KAN - I is NPF -8 7KAN
- IA and NPF-87KAN-rB are about a 1:6 mixture, while NPF-87
KAN-II NPF 8 7KAN-II mA
and about l to 4 of NPF 8 7KAN-II B
It was recognized that it was a mixture of

As will be discussed later, regarding the growth inhibition activity of dental caries bacteria, NPF
8 7KAN-IA, NPF-87K
AN-IB, NPF-8 7KAN-I
IA and NPF-87KAN-IIB and NPF
- 8 7KAN -■ and NPF - 8 7KAN
- I[ has almost the same effect.

The effective substance is soluble in methanol, ethanol, and water, so the extraction method described above starts from a methanol extract as a raw material, but in order to save expensive organic solvents, a large amount of water is first needed. Alternatively, the same operation may be performed after extraction with hot water.

Furthermore, as is clear from the ultraviolet absorption spectra mentioned above, when made alkaline, the new substances of the present invention change color from yellowish brown to reddish brown, so the entire extraction process is carried out under weakly acidic conditions using mineral acids or organic acids. This is also an effective extraction tool.

Even a crude extract with methanol such as methanol has the activity of inhibiting the growth of dental caries bacteria and can be used as an active ingredient in the caries preventive agent of the present invention. It is preferable to use this method.

■． The effective substance of the present invention obtained from the caries preventive agent is a polyphenolic compound. Conventionally, certain polyphenolic compounds have been known to inhibit adhesive glucan production or enzymes (glucosyltransferase), for example, as disclosed in JP-A-58-121218. , there is no mention of polyphenols in Kanshu.

In addition, it has recently been reported that low-molecular polyphenols obtained from Sencha green tea have the effect of inhibiting the growth of caries bacteria (Japanese Patent Application Laid-Open No. 64-90124). It is.

The caries preventive agent of the present invention is obtained by extracting the active ingredient by various methods such as extracting the caries with methanol, or NPF 8 7KAN-I, NPF-8 7
KAN-I [, NPF 8 7KAN-I
A, NPF-87KAN-IB, NPF-87KAN
-IfA and NPF - 8 7KAN -II B
It is produced by adding it to a known oral cavity composition in the form of a purified product.

Known oral compositions include toothpaste, toothpaste, mouthwash, candies, chewing gum, various sweets, all other beverages, foods, and oral medications.

■． When acute toxicity was determined by intraperitoneal administration to mice, the following values were obtained.

NPF 87KAN I LDso=200
mg/kgNPF-8 7 KAN-n
L Dso >400 mg/kgNPF
87K,AN-IA LDso=200mg/k
gNPF-8 7KAN-IBLD
so=200 mg/kgNPF 87KAN
-ILA LDso>400mg/kgNPF
8 7 KAN II BL Dsa >40
0 mg/kg [Example] Hereinafter, the present invention will be explained in more detail with reference to Examples and Reference Examples.

Reference example 1: Extraction of active substances a) Crushed and dried Kanshu [Seller: Uchida Japanese and Chinese Medicine] 5
00g to 1,500mj of hexane! After immersing it in the solution and leaving it at room temperature for 24 hours, hexane was removed by filtration. This operation was repeated three times to remove fat.

The yield of hexane soluble portion was 1.11 g.

b) The defatted kanshu was air-dried for one day.

c) Pour the air-dried ingredients into 1.0 ml of methanol. 000 and extracted under boiling for 3 hours. Do this simulation three times,
The extract was collected.

2) Collect the obtained extract and use an evaporator in water bath 4.
It was condensed under reduced pressure at 0°C and dried under vacuum (yield: i.5 g).
.

500 g of water was added to this, stirred, and then filtered.

The insoluble matter is an additional 200mj of water! was added and stirred, followed by filtration and the filtrate was collected.

E> 500 ml of ethyl acetate was added to this aqueous solution for extraction. This operation was repeated three times, and the ethyl acetate soluble portion H. 0
37) was removed. 1.037 ml of ethyl acetate extract was obtained.

f) The water extract from which the ethyl acetate soluble portion has been removed has a molecular weight of 1. 0 0 0 dialysis tube (Svectra/bore 6; manufactured by Svectrum Medical Industry Co., Ltd.) and stirred with water at 4°C for 1 week, the external solution was exchanged every day during dialysis to separate the internal and external solutions. It was fractionated into a liquid.

g) The dialysis fluid fractionated with a dialysis tube with a molecular weight cutoff of 1 and OOOO is further put into a dialysis tube with a molecular weight cutoff of 10,000 (Svectra/Bore 6; Svectra Medical Industry Co., Ltd.!!), Dialyze with water at 4°C for 1 week while stirring, changing the external solution daily during that time.
Fractionate into internal and external fluid.

h) When fractionated in this way, the molecules it. 0 0
The desired activity of inhibiting the growth of caries bacteria was observed in the fractions above 0 to 10,000 and 10,000, and by freeze-drying,
2 active substances! Both OA could be obtained as light brown powder. Molecule I1. 0 0 0 to 10,000 fraction (NPF 8 7KAN-I) was 1. 7
3 g of the fraction with a molecular weight of 10,000 or more (NPF 8 7KAN-II) was 1 1.
I was able to get 8g.

Molecular weight 1. The fraction below 0 0 0 is 7. 9 2
However, the growth inhibition activity of caries bacteria was higher than that of NPF.
-87KAN-i and NPF 8 7KAN-
It was very weak compared to If.

Reference example 2: Extraction of effective amount a) Crushed and dried Kanshu [Sold by 21 Kinokuniya Kan Pharmacy]
50 g was immersed in 150 grni of hexane, left at room temperature for 24 hours, and then the hexane was removed by filtration. This operation was repeated three times to remove fat. The yield of hexane soluble portion is 0. It was 10g.

Mouth) The defatted husks were air-dried for 1 day.

c) The air-dried husks were immersed in 100 rnl of methanol and extracted under boiling for 3 hours. This operation was performed three times and the extracts were collected.

2) Collect the obtained extract and use an evaporator in a water bath for 40 minutes.
It was condensed under reduced pressure at °C and dried under vacuum.

(Yield: 2.07 g) The yield of the extract in the above steps is as shown in Table 1.

Table 1 Reference Example 3 NPF-8 7KAN-1 and NPF 87
KAN If was subjected to high performance liquid chromatography and N
PF-8 7KAN-IASNPF-8 7
KAN-IBSNPF-8 7KAN-IIA
and NPF-8 7KAN-IIB were isolated. The conditions are as follows.

Separation power ram: Packed with reverse phase distribution type packing material (TSKg
el ODS-8 0TIvf: manufactured by Tosoh ■) Eluent nimethanol (contains 1% acetic acid) Detector: Ultraviolet spectroscopic detector (manufactured by JASCO Corporation) 28 Or + m NPF-87KAN -I 500 ■ NPF
- 8 7 KANIA 49.5mg NPF-
80 mg of 87KAN I B2 was obtained.

In addition, NPF 87KAN ffA'37.1mg than NPF 87KAN I [250mg
NPF 87KAN-I [B 1 8 4■ was obtained.

Example 1: Effect of inhibiting growth of dental caries bacteria (1) (method) (a) Subcultured and grown dental caries bacteria (S, mutans)
, MT8148(C), S. mutans 671.
5 Add sterilized physiological saline 8- to the slant in (1) to obtain a bacterial suspension.

(b) RPMI l, 5% agar added l 6 4
0 (pH 7.0) C Nissui Pharmaceutical ■], NPF
-87KAN -1 and ~PF -8 7KAN -I
I and as a control molecular weight cutoff 1, O O O or more 1
, 0 0 0 or less, prepare 10 ml of culture medium for each concentration of Japanese tea water extract, 8. The mixture was placed in a 5 cm petri dish to prepare an agar plate.

(c) 100μ of the suspension prepared in (c) on this agar plate
The minimum growth inhibitory concentration (abbreviated as MIC) was determined by uniformly applying 1 and culturing at 30° C. overnight.

Incidentally, if the growth status of the plants has not yet been determined, after culturing them overnight, they are grown in agar-free medium 5rnI. was added, cultured, and MIC was determined.

The above results are shown in Table 2.

Example 2: Effect of inhibiting growth of dental caries bacteria (2) (Method) (a) The following two types of culture solutions for dental caries bacteria, which were the same as those used in Example 1, were prepared.

A medium: ASF medium 104 (Ajinomoto ■) with 1% fetal bovine blood added, B medium + ASF medium 104 with 1% glucose added, (+1) Dental caries bacteria in A medium overnight at 30°C. The cells were cultured and made into a bacterial solution.

(C) 8 ml of culture medium with each concentration of the specimen was prepared using 8 and B medium, 100 mj' of (+1) bacterial solution was added thereto, and cultured overnight at 30°C to determine the MIC.

Judgment was made with the naked eye or by absorbance at 660 nm.

The results are shown in Table 3.

[Brief explanation of drawings]

FIG. 1 shows the infrared spectrum of NPF-87KAN-1. FIG. 2 shows the ultraviolet absorption spectra of NPF-87KAN-I in water and 0.1N hydrochloric acid solutions. Figure 3 shows 0.1 of NPF-87 KAN-I.
The ultraviolet absorption spectrum of normal sodium hydroxide solution is shown. FIG. 4 shows the infrared absorption spectrum of NPF 8 7KAN-II. FIG. 5 shows the ultraviolet absorption spectra of NPF-87KAN-n in water and 0.1N hydrochloric acid solutions. Figure 6 shows the NPF. - 8 7KAN - If's 0.
The ultraviolet absorption spectrum of a 1N sodium hydroxide solution is shown. FIG. 7 shows the infrared absorption spectrum of NPF-8 7KAN-IA. FIG. 8 shows the infrared absorption spectrum of NPF-87KAN-IB. FIG. 9 shows the infrared absorption spectrum of NPF-8 7KAN-IrA. FIG. 10 shows the infrared absorption spectrum of NPF-87KAN-IIB. Figure 11 A and B are NPF-8 7KAN-
1 and NPF-8 7KAN-II are shown. (A) (NPF-87KAN-1) NPF-87KAN-IA (B) (NPF-87KAN-II) NPF-87KAN-mA

Claims (3)

[Claims] (1) NPF-87KAN-I, NPF-87KAN-II, N having the following physical and chemical properties
PF-87KAN-IA, NPF-8KAN-IB,
NPF-87KAN-IIA and NPF-87KAN-I
A caries prevention agent whose active ingredient is a substance selected from the group consisting of IB. (NPF-87KAN-I) (i) Shape: Light brown powder (ii) Melting point: No clear melting point or decomposition point. (iii) Elemental analysis: C: 49.14% H: 5.42% N: <0.5% Ash: <0.9% (iv) Molecular weight: 1,000-10,000 (by dialysis tube) (v) Infrared absorption spectrum: ν_m_a_xcm^-^1; 3394, 2938, 1
617, 1521, 1443, 1375, 1284, 1
245, 1103, 1067, 779 (vi) Ultraviolet absorption spectrum: λ^Water_m_a_x nm (E^1^%_1_c_m)
280 (74.2) λ^0^-^1_m_a_x Normal hydrochloric acid nm (E^1^%_1_c_m) 280 (73.
4) λ^0^-^1_m_a_x Normal sodium hydroxide nm (E^1^%_1_c_m) 305 shoulder (10
7.7) 510 shoulder (21.6) (vii) Solubility: Soluble in water, methanol, ethanol. Insoluble in hexane, ether, ethyl acetate, and chloroform. (viii) Color reaction: Ferric chloride Positive ninhydrin Negative p-anisidine-phthalic acid Anisidine-diphenylamine Dragendorff (NPE-87KAN-II) (i) Shape: Light brown powder (ii) Melting point: Clear It shows no melting point or decomposition point. (iii) Elemental analysis: C: 53.58% H: 4.55% N: <0.3% Ash: <0.3% (iv) Molecular weight: 10,000 or more (by dialysis tube) (v) Infrared absorption spectrum: ν_m_a_xcm^-^1; 3422, 2926, 1
609, 1518, 1441, 1392, 1286, 1
241, 1103, 1063, 789 (vi) Ultraviolet absorption spectrum: λ^Water_m_a_x nm (E^1^%_1_c_m)
280 (129.4) λ^0^-^1_m_a_x Normal hydrochloric acid nm (E^1^%_1_c_m) 280 (12
8.1) λ_m_a_x normal sodium hydroxide nm
(E^1^%_1_c_m) 510 shoulders (25.5) (v
ii) Solubility: Soluble in water, methanol, ethanol. Insoluble in hexane, ether, ethyl acetate, and chloroform. (viii) Color reaction: Ferric chloride Positive ninhydrin Negative p-anisidine-phthalic acid Anisidine-diphenylamine Dragendorff (NPF-87KAN-IA) (i) Shape: Light brown powder (ii) Melting point: Clear It shows no melting point or decomposition point. (iii) Elemental analysis: C: 37.71% H: 3.97% N: 2.83% Ash: 16.48% (iv) Molecular weight: 8,100 (by gel permeation chromatography based on polyethylene glycol) ) (v) Infrared absorption spectrum: ν^k^a^r_m_a_xcm^-^1; 3426,
2920, 2852, 1620, 1445, 1384,
1259, 1114, 1067, 802 (vi) Ultraviolet absorption spectrum: λ^Water_m_a_x nm (E^1^%_1_c_m)
280(vii) Solubility: Soluble in water, methanol, ethanol. Insoluble in hexane, ether, ethyl acetate, and chloroform. (viii) Color reaction: Ferric chloride Positive ninhydrin Negative p-anisidine-phthalic acid Negative anisidine-diphenylamine Negative Dragendorff negative (NPF-87KAN-IB) (i) Shape: Light brown powder (ii) Melting point: Clear It shows no melting point or decomposition point. (iii) Elemental analysis: C: 45.78% H: 4.45% N: 0.71% Ash: 10.45% (iv) Molecular weight: 4,900 (by gel permeation chromatography based on polyethylene glycol) ) (v) Infrared absorption spectrum: ν^k^a^r_m_a_xcm^-^1; 3410,
2964, 1609, 15271442, 1284, 1
262, 1206, 1157, 1113, 1067, 8
03 (vi) Ultraviolet absorption spectrum: λ^Water_m_a_x nm (E^1^%_1_c_m)
280(vii) Solubility: Soluble in water, methanol, ethanol. Insoluble in hexane, ether, ethyl acetate, and chloroform. (viii) Color reaction: Ferric chloride Positive ninhydrin Negative p-anisidine-phthalic acid Negative anisidine-diphenylamine Negative Dragendorff negative (NPF-87KAN-IIA) (i) Shape: Light brown powder (ii) Melting point: Clear It shows no melting point or decomposition point. (iii) Elemental analysis: C: 38.66% H: 3.67% N: 1.58% Ash: 22.94% (iv) Molecular weight: 12,800 (by gel permeation chromatography based on polyethylene glycol) ) (v) Infrared absorption spectrum: ν^k^b^r_m_a_xcm^-^1; 3402,
1611, 1521, 1442, 1372, 1283,
1205, 1113, 795 (vi) Ultraviolet absorption spectrum: λ^Water_m_a_x nm (E^1^%_1_c_m)
280(vii) Solubility: Soluble in water, methanol, ethanol. Insoluble in hexane, ether, ethyl acetate, and chloroform. (viii) Color reaction: Ferric chloride Positive ninhydrin Negative p-anisidine-phthalic acid Negative anisidine-diphenylamine Negative Dragendorff negative (NPF-87KAN-IIB) (i) Shape: Light brown powder (ii) Melting point: Clear It shows no melting point or decomposition point. (iii) Elemental analysis: C: 44.97% H: 4.44% N: 0.79% Ash: 14.32% (iv) Molecular weight: 9,400 (by gel permeation chromatography based on polyethylene glycol) ) (v) Infrared absorption spectrum: ν^k^a^r_m_a_xcm^-^1; 3388,
2964, 1611, 1519, 1445, 1379,
1285, 1261, 1202, 1096, 802 (vi) Ultraviolet absorption spectrum: λ^Water_m_a_x nm (E^1^%_1_c_m)
280(vii) Solubility: Soluble in water, methanol, ethanol. Insoluble in hexane, ether, ethyl acetate, and chloroform. (viii) Color reaction: Ferric chloride Positive ninhydrin Negative p-anisidine-phthalic acid Negative Anisidine-diphenylamine Negative Dragendorff Negative (2) The molecular weight of the defatted methanol extract obtained by fractionating it using a dialysis tube is 1,000 to 10,0.
An anti-caries agent containing a hydrophilic solvent extract of the 0.00 fraction as an active ingredient. (3) An anti-caries agent containing as an active ingredient a hydrophilic solvent extract of delipidum, which is a fraction with a molecular weight of 10,000 or more obtained by fractionating a defatted methanol extract of delipidum using a dialysis tube. (4) A caries preventive agent containing a hydrophilic solvent extract of Kanshu as an active ingredient.