JP2002154959A - Analgesic and anti-inflammatory agent - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an analgesic and anti-inflammatory agent having both high analgesic and anti-inflammatory activities substantially associated with no side effect. SOLUTION: This analgesic and anti-inflammatory agent comprises as the active ingredient a pharmaceutically acceptable water-soluble salt of incarvillateine of formula (I); wherein the compound of formula (I) is preferably such as to be extracted from Incarvillea sinensis LAM in particular. This agent is characterized by being drug-nondependent.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an analgesic and anti-inflammatory agent. More specifically, the present invention relates to an analgesic and anti-inflammatory agent containing a water-soluble salt of a compound extracted from a plant as an active ingredient.

[0002] The various diseases with the background technology severe pain, for example terminal cancer, in, analgesic or relief of cancer pain has become an important nursing issues. To date, a number of compounds have been reported that exhibit analgesic and anti-inflammatory effects on cancer pain. Plants such as poppy are generally well known as crude drugs having such an action. Poppies include opium, the main components of which are morphine and morphine derivatives, which are known to have high analgesic, antispasmodic, antitussive and antidiarrheal effects. However, morphine has the problem of increased drug dependence when used continuously, and may further have side effects such as constipation and respiratory depression. For this reason, morphine is designated as a drug because of its strong dependence on drugs, and its use is often restricted because of its strict management requirements. Therefore, an analgesic having no side effects such as drug dependence has been desired.

The natural plant Incarvillea sinens
is LAM. ) is a plant belonging to the family Bignoniaceae, and is known and widely used as a crude drug having a detoxifying effect, an anti-rheumatic effect, and an analgesic effect in China. It is also known that a crude drug using kakuko hardly causes a problem of side effects associated with its use. For this reason, it is expected that an ingredient containing gongko may be used as an analgesic and anti-inflammatory agent. Recently, it has been revealed that the main analgesic activity contained in Susumu is Incarvillateine, a kind of monoterpene alkaloid [(1) Yu-Ming Chi, Wen-Mei Yan, a
nd Jia-Shi Li: Phytochemistry. Vol.29, No.7, pp2
376-2378, 1990, and (2) M. Nakamura, Yu-Ming C
hi, Wen-Mei Yan, Y. Nakasugi, T. Yoshizawa, N. Iri
no, F. Hashimoto, J. Kinjo, T. Nohara, and S. Saku
rada: J. Nat. Prod., Vol.62, pp1293-1294, 1999
Year].

[0004] However, if the incabiratein extracted from Kakuho is used as it is, the expected effect may not be sufficiently exhibited depending on the administration route. In some cases, it is difficult to form a desired dosage form from the above. For this reason, when using incabiraatein as an analgesic, its application range was often restricted.

[0005]

SUMMARY OF THE INVENTION The present inventors have recently formed a water-soluble salt using incabiraatein extracted from Ganko,
It has been found that by using it, an excellent analgesic and anti-inflammatory effect can be obtained with almost no side effects as seen in conventional narcotic analgesics such as morphine. The present invention is based on this finding. Therefore, an object of the present invention is to provide an analgesic and anti-inflammatory agent excellent in analgesic action and anti-inflammatory action with almost no side effects.

The analgesic and anti-inflammatory agent according to the present invention comprises a pharmaceutically acceptable water-soluble salt of incabiraate as an active ingredient.

[0007] In the present invention, since a component derived from Ganko, which is conventionally used as a crude drug and is known to hardly cause side effects such as dependence, is used, the analgesic and anti-inflammatory according to the present invention is used. An agent is one that has few side effects. A pharmaceutically acceptable water-soluble salt of incabiraate, which is an active ingredient of this analgesic and anti-inflammatory agent, has the same action as an analgesic action of an opioid-type narcotic analgesic such as morphine. (Single-phase reaction), but can exert a more powerful analgesic and anti-inflammatory effect than in the case of incabiratein. Inkaviratein is a low-polarity substance and is insoluble in water.However, since the water-soluble salt of the incaviratein is water-soluble, unlike incaviratein, it has a broader range than incaviratein. It can be used in any route of administration and can show excellent analgesic and anti-inflammatory effects at lower doses. In addition, the analgesic anti-inflammatory agent according to the present invention also exhibits an anti-inflammatory effect (two-phase reaction) as shown by ordinary analgesics such as aspirin, so that it can be used not only for cancer pain and the like, but also for headache, abdominal pain, Also, excellent effects can be exerted on neuralgia and the like.

[0008]

DETAILED DESCRIPTION OF THE INVENTION Analgesic and anti-inflammatory agent The analgesic and anti-inflammatory agent according to the present invention comprises a pharmaceutically acceptable water-soluble salt of incabillatein as an active ingredient. Here, "comprising as an active ingredient" naturally includes the case where a carrier corresponding to the dosage form is included, and also means that other drug components that can be used in combination may be contained. As such a drug component that can be used in combination, for example, a component of a conventional drug having an analgesic effect such as morphine, aspirin, indomethacin, or the like, or a drug component for the purpose of gastrointestinal protection, etc. Is exemplified. Furthermore, examples of such a drug component that can be used in combination include components such as a preservative, a binder, a stabilizer, a lubricant, and a flavor.

[0009] Inkaviratein is a compound represented by the following formula (I) and can be obtained from natural sources.
For example, Inkabilatein is contained in the plant of Incarvillea sinensis LAM., Which is a plant of the family Ceratopogonaceae, and can be obtained from here. In the present invention, incaviratein is not particularly limited to those derived from Kakuko, and may be derived from other plants, but is preferably extracted from Kakuko.

[0010]

Embedded image

[0011] The extraction of incabiraatein from Susumu can be carried out as follows, for example.
That is, the whole plant of corn plant is usually used for the extraction of incabiraatein. This can be raw or dried,
Crushed and then water (preferably hot water) as extraction solvent
Alternatively, extraction is carried out using an organic solvent, and if necessary, a combination of an organic solvent and water. As the organic solvent in the extraction, for example, methanol, ethanol, n-
Butanol or ethyl acetate is used, preferably methanol or ethanol. The extraction operation is usually performed under room temperature conditions or reflux conditions. The extract thus obtained is subjected to chromatography using, for example, a polystyrene column, and a component having a remarkable analgesic and anti-inflammatory effect is separated from the obtained fraction,
By further purifying this component, incabiraatein can be obtained.

In the present invention, the “pharmaceutically acceptable water-soluble salt of incabiraatein” is basically any salt of incabiraatein as long as it is soluble in water. Are also included. Therefore, for example, it may be a salt such as an acidic salt or a basic salt, or may be a salt such as a double salt or a complex salt. Furthermore, in the present invention, hydrates of the above-mentioned salts are also included. In a preferred embodiment of the present invention, as the pharmaceutically acceptable water-soluble salt of incabillatein, an inorganic acid salt of incabillatein or an organic acid of incabillatein is preferably mentioned.

The above-mentioned inorganic acid salts include hydrochloric acid, sulfuric acid,
A preferred example is a salt of an inorganic acid such as hydrobromic acid, nitric acid, phosphoric acid, or carbonic acid and incabiraatein. Further, as the above-mentioned organic acid salt, acetic acid,
Salts of organic acids such as benzoic acid, oxalic acid, succinic acid, tartaric acid, citric acid, maleic acid, fumaric acid, malic acid, sulfonic acid, sulfinic acid, or propionic acid and incabiraatein are preferred. Can be.

The general formula of a pharmaceutically acceptable water-soluble salt of incabiraate preferably used in the present invention can be represented by, for example, the following formula (II). C ₄₂ H ₅₈ N ₂ O ₈ R (II) wherein R is 2HCl, H ₂ SO ₄ , 2HBr, 2H
NO ₃ , 2CH ₃ COOH, or 2C ₆ H ₅ COOH
Can be ]

Of these, for example, the structural formula of incabiraate hydrochloride can be represented by the following formula (III).

Embedded image

In the present invention, a pharmaceutically acceptable water-soluble salt of incabiraate can be obtained, for example, by adding an inorganic acid or an organic acid to incabiraate obtained by extracting and purifying from inocula. It can be obtained by performing heat treatment accordingly. More specifically, for example, the obtained incabiraatein is added to an alcohol solution of an inorganic acid or an organic acid, and after heating and reacting, the reaction solution is mixed with a large amount of a low-polarity solvent (but mixed with alcohol). Things) (eg, ether, chloroform, benzene, etc.)
The water-soluble salt of incabiraatein can be obtained by injecting the mixture into the flask and washing the precipitate that precipitates several times. This salt can be easily ionized and dissolved in an aqueous solution. Furthermore, if a specific example of the method for producing the water-soluble salt is given, it can be produced by the following procedure. That is, incabilatein is replaced with 2N-HC
1 / MeOH and heated at 60 ° C. for 1 hour to react. After completion of the reaction, the reaction solution is poured into a large amount of ether, and the deposited precipitate is collected by filtration, dried and washed several times with ether. Incaviratein hydrochloride can be obtained as a white amorphous powder.

The pharmaceutically acceptable water-soluble salt of incabiraate used in the analgesic and anti-inflammatory agent according to the present invention includes:
As described above, those that do not show drug dependence such as chronic intoxication symptoms, that is, those that are drug-independent. The analgesic and anti-inflammatory agent according to the present invention may comprise, in addition to the water-soluble salt of incabiraate, other concomitant drug components and the like. It is preferable that the agent as a whole does not show drug dependence, that is, it is drug-independent.

The analgesic and anti-inflammatory agent according to the present invention is usually provided in the form of a pharmaceutical or food. When used as a pharmaceutical, the analgesic and anti-inflammatory agent according to the present invention can be provided in the form of a dry powder, powder, granule, tablet, dragee, capsule, wrap, drop, etc. Since the salt of incabiraate used as a component is water-soluble, the analgesic and anti-inflammatory agent can be provided in the form of a liquid or a liquid-encapsulated capsule. Further, the analgesic and anti-inflammatory agent according to the present invention may be used after dissolving a dry powder or the like in water.
At this time, the analgesic anti-inflammatory agent, depending on the intended dosage form and conditions of use, other components, for example, excipients, preservatives,
It can include binders, stabilizers, antistatic agents, lubricants, flavors, and the like. When used as food, the analgesic and anti-inflammatory agent according to the present invention can be provided in the form of gum, candy, jelly, tablet confectionery, beverage and the like.

When administered as a pharmaceutical, the analgesic and anti-inflammatory agent according to the present invention can adopt various administration forms such as oral administration, parenteral administration, inhalation, rectal administration and topical administration. Among these, parenteral administration includes subcutaneous injection, intravenous administration, intramuscular administration, intranasal administration or infusion. Therefore, for example, mucosal administration, transdermal administration, or administration with an implant for the nose, mouth, sublingual, rectum and the like is also possible. For example, when the analgesic and anti-inflammatory agent according to the present invention is provided in the form of a dry powder, which is to be used as an injection, the powder is mixed with distilled water or sodium chloride and saccharides (glucose, mannitol, inositol, etc.). After dissolving in the prepared isotonic aqueous solution, the active ingredient is a drug concentration in vivo that is effective against the target disease, and is used as an injection for intravenous, intramuscular, subcutaneous, intraorgan, or direct tumor or tumor resection Can be administered to the lesion.

The dose of the analgesic anti-inflammatory agent according to the present invention is
In terms of the water-soluble salt of incabiraate, it is generally in the range of about 0.1 to 500 mg / kg body weight per dose, preferably in the range of 0.1 to 200 mg / kg body weight,
Usually, it is preferably administered 1 to 5 times a day. However, the exact dose is preferably determined within the above range in consideration of the patient's age, weight, condition, administration route, and the like. The toxicity of the analgesic anti-inflammatory agent according to the present invention is low. That is, when the acute toxicity when the analgesic and anti-inflammatory agent was orally administered was examined using male Wistar rats, at least 3000 m
It has been confirmed that no deaths occurred in rats up to the dose (dose) of g / kg (po).

[0021]

DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will be described in detail below with reference to examples, but the present invention is not limited to these examples.

Preparation of Sample Preparation of Sample 1
The residue was extracted with 5.5 L of methanol at room temperature, and the extracted methanol solution was concentrated under reduced pressure to obtain 42.7 g of a dark green powdery extract (sample 1). ) Got.

Preparation of Samples 2 to 8 Sample 1 was subjected to column chromatography using 230 ml of a polystyrene resin (trade name: Diaion HP-20, styrene-divinylbenzene copolymer resin, manufactured by Mitsubishi Chemical Corporation). And 10% or 30% methanol in 2 L each, 40%, 50%, 60%, 70%
% Or 80% methanol was eluted with 3 L each, and each eluate was concentrated under reduced pressure to obtain the following Samples 2 to 8, respectively. Sample 2: 10% methanol extraction fraction 1.7
g Sample 3: 30% methanol extraction fraction 0.6
g Sample 4: 40% methanol extraction fraction 1.0
g Sample 5: 50% methanol extraction fraction 1.4
g Sample 6: 60% methanol extraction fraction 1.5
g Sample 7: 70% methanol extraction fraction 1.9
g Sample 8: 80% methanol extraction fraction 0.7
g

Preparation of Sample 9 The sample 6 (280.3 mg) was mixed with Sephadex L
Column chromatography using 500 ml of H-20 (manufactured by Pharmacia LKB Biotechnology, polysaccharide dextran beads) was performed, and eluted with 30% methanol. The eluate was fractionated by 5 ml each to obtain fraction 1
~ 20 were obtained. Among these, 71.7 mg of a colorless powder was obtained from fractions 8 to 10. The obtained colorless powder was repeatedly recrystallized using methanol and purified to obtain colorless plate-like crystals (Sample 9). The physicochemical properties of the obtained sample 9 were examined by melting point measurement, mass spectrometry, nuclear magnetic resonance (NMR), and the like. The results were as shown below.

M. p. 217.2-217.7 ° C [a] _D -10.8 ° (c = 0.79, CHCl ₃ ) EI-MS (m / z): 718 [M] ^{+ 13} C-NMR (CDCl ₃ -d ) d: 171.9 (s), 17
1.7 (s), 146.9 (s), 146.8 (s), 145.5 (s), 145.3 (s), 13
0.4 (s), 130.2 (s), 120.3 (d), 119.8 (d), 114.7 (d), 11
4.7 (d), 110.9 (d), 110.8 (d), 76.5 (d), 76.2 (d), 57.4
(t), 57.3 (t), 57.1 (t), 57.1 (t), 55.7 (q), 55.6 (q),
47.8 (d), 47.2 (d), 46.0 (q), 45.9 (q), 45.7 (s), 45.6
(s), 41.7 (d), 40.3 (d), 40.1 (d), 40.1 (d), 37.2 (s), 3
7.2 (s), 30.1 (d), 30.1 (d), 29.6 (t), 29.1 (t), 17.0
(q), 16.8 (q), 14.8 (q), 14.4 (q)

The above results were reported in the literature (Yu-Ming Chi, Wen-M
ei Yan, Jia-Shi Li: Phytochemistry. Vol.29, No.7,
pp. 2376-2378, 1990), it was confirmed that this sample 9 was incabiralatein by comparing with literature values of incabiraatein isolated from the plant described in the present invention.

Preparation of Sample 10 The sample 9 (40 mg) was treated with 2N HCl / MeOH (5
mL) and heated at 60 ° C. for 1 hour to allow the reaction to proceed. After completion of the reaction, the obtained reaction solution was poured into a large amount of ether, and the deposited precipitate was collected by filtration, dried and washed several times with ether to obtain a white amorphous powder (sample 10).
I got The yield was 73.0%. The physicochemical properties of the obtained sample 9 were examined by mass spectrometry, nuclear magnetic resonance (NMR), and the like. The results were as shown below.

[A] _D -8.2 ° (c = 0.80, MeOH) HR-FAB MS m / z: 755.4036 [M] ⁺ calcd. For C ₄₂ H
₆₀ N ₂ O ₈ Cl: 756.37 ¹³ C-NMR (CDCl ₃ -d ) d: 172.0 (s), 1
71.8 (s), 147.0 (s), 147.0 (s), 145.4 (s), 145.2 (s), 1
30.4 (s), 130.4 (s), 120.4 (d), 119.9 (d), 114.9 (d), 1
14.9 (d), 111.2 (d), 111.2 (d), 75.5 (d), 75.2 (d), 56.
1 (q), 56.1 (q), 55.2 (t), 55.2 (t), 54.4. (T), 54.4
(t), 48.0 (d), 47.1 (d), 43.6 (q), 43.6 (q), 43.1 (s), 4
2.9 (s), 41.8 (d), 41.2 (d), 40.5 (d), 40.5 (d), 36.2
(s), 36.1 (s), 29.7 (t), 29.1 (t), 28.3 (d), 28.2 (d), 1
6.3 (q), 16.0 (q), 14.5 (q), 14.1 (q)

From the above results, it was confirmed that this sample 10 was hydrochloride of incabiraatein.

Evaluation Test (1) Analgesic Anti-Inflammatory Activity Test An analgesic anti-inflammatory activity test (so-called formalin test) was performed using Samples 9 and 10 as test drugs. 20 μl of 0.2% formalin solution as a stimulant mouse (purchased for one week after purchasing a 5-week-old male ICR strain)
After injection into the hind leg paw (subcutaneously on the sole of the foot), the licking action of the mouse on the hind leg to alleviate pain was measured using a stopwatch for 5 consecutive minutes for 30 minutes. At this time, for each test drug (samples 9 and 10), 0.5% was set so that the dose of the nuclear test drug was equivalent to 10 mg / kg.
% Aqueous solution of polyoxyethylene sorbitan monooleate (trade name: Tween 80, manufactured by Atlas Co.)
100 μl of each suspension were prepared. These were administered to mice intraperitoneally 15 minutes before stimulation. As a control group (control group), 0.5%
The same volume of Tween 80 was administered intraperitoneally to mice.

It is generally known that when such a formalin test is performed, the following two reactions occur, namely, a one-phase reaction (First Phase) and a two-phase reaction (Second Phase). The monophasic reaction refers to a response due to pain between 0 and 10 minutes after formalin stimulation, and is a response due to pain caused by directly stimulating the terminals of sensory nerves by injecting formalin. At present, only narcotic analgesics such as morphine have an action of suppressing this, and its mechanism of action is considered to be acting on the central nervous system. On the other hand, the biphasic reaction is
It refers to a response due to pain during 10 to 30 minutes, and refers to a response due to pain when inflammation is caused by injecting formalin. This reaction can usually be suppressed with analgesics such as aspirin.

The evaluation method of the analgesic anti-inflammatory activity test is from 0 to 1
The evaluation was performed by dividing into two phases: a phase until 0 minutes (one phase) and a phase from 10 to 30 minutes (two phases). In each case, the time the mouse licked its hind legs (Licking Tim
e) was calculated as the total number of seconds and the value was compared with the control.

The results obtained (n = 10, mean ± standard error) are as shown in Table 1 and FIG.

Table 1 : Results of formalin test [Test conditions: When stimulated with 1% formalin (injection of 20 ml), intraperitoneal administration (10 animals per group)] One-phase reaction (0 to 10 minutes) Two-phase reaction (10 to 30 minutes) Min) control 131.46 ± 7.84 164.50 ± 19.68 Sample 9 71.21 ± 7.16 ** 6.11 ± 1.69 ** Sample 10 18.33 ± 3.80 ** 10.61 ± 2.74 ** ** P <0.01

(2) Location Preference Test (Conditioned Plac
e Preference test) Whether or not the sample 10 caused drug dependence as a side effect was examined by measuring the presence or absence of the formation of mental dependence by a place preference test as described below. In addition, it is known that an opioid analgesic represented by morphine forms dependence as a side effect. In the place preference test, specifically, after preconditioning the relationship between the effect of the drug and the sight (white and black) of the two black and white compartment boxes on the mouse, the mouse was allowed to select these two boxes, When a mouse prefers a drug-treated box, the drug is evaluated as having a rewarding effect (mental dependence), and conversely, when the mouse prefers a solvent-treated box, it is evaluated as having an aversive effect.

As test subject mice, 4-week-old male ddY strain (25 ± 5 g) mice that had been quarantined for 1 week after purchase were used, and 10 mice were included in one group. For the control group, 0.5% Tween 80 physiological saline 100 μm
1 and 0.5% for the morphine group
Morphine 4mg / k in Tween 80 saline
g or 8 mg / kg of the morphine suspension adjusted to correspond to a dose of 8 mg / kg, and for group 10 samples, 20 mg / kg or 40 mg / kg in 0.5% Tween 80 physiological saline as well. These were administered to mice (subcutaneously) using 100 ml of the 10 suspensions of the sample adjusted to correspond to the doses of.

The mouse to which the test drug or control solvent was administered was confined in one box and left for one day.
Conversely, the drug administration group on the previous day (morphine group and sample 10)
Group) received control solvent and was confined in the other box for 1 hour. This operation was repeated twice (4 days), and the relationship between the effect of the drug and the sight of the two compartment boxes was conditioned on the mice. Then, on the fifth day, the central partition of both boxes was removed, and the platform was installed. The conditioned mice were placed on the platform, and the time spent in each compartment for 15 minutes after landing on the floor was measured to evaluate the reward effect and the aversion effect. Evaluation was performed by comparing each drug administration group with a control group according to Dunnett's multiple comparison test method. It should be noted that the value of the place preference represents the average value of the time spent in one certain box,
At this time, the staying time of the box on the drug-treated side was indicated by plus, and the staying time of the box on the solvent side (non-drug-treated side) was indicated by minus. That is, the value of the place preference is
The plus side indicates that there is a tendency to reward, and the minus side indicates that there is a tendency to dislike.

Obtained result (place preference) (n = 10,
The average value ± standard error) was as shown in Table 2 and FIG.

Table 2 : Results of place preference test [Test conditions: morphine and sample 10: subcutaneous administration (10 animals per group)] Administered drug Place preference (sec) control -35.20 ± 45.30 Morphine (4 mg / kg) 74.50 ± 60.90 Morphine (8 mg / kg) 203.50 ± 42.10 Sample 10 (20 mg / kg) -53.70 ± 59.20 Sample 10 (40 mg / kg) -96.40 ± 49.80

[Brief description of the drawings]

FIG. 1 is a diagram showing the results of an analgesic and anti-inflammatory activity test (formalin test) by intraperitoneally administering a control, sample 9 and sample 10 to mice, respectively.
rst Phase = 0 to 10 minutes, Second Phase = 10 to 30
Minutes).

FIG. 2. Control, morphine, and sample 10
It is a figure which shows the result of the place preference test (Conditioned Place Preference test) by subcutaneously administering to each mouse.

 ────────────────────────────────────────────────── ─── Continuing on the front page (72) Inventor Minoru Nohara Hiroshi 2-41-4 Nagaminehigashi, Kumamoto City, Kumamoto Prefecture (72) Inventor Shinobu Sakurada 7-3-1 Takamori, Izumi-ku, Sendai City, Miyagi Prefecture F-term ( Reference) 4C034 CB01 4C086 AA01 AA02 BC27 GA07 MA01 MA04 NA14 ZA08 ZB11

Claims (4)

[Claims] An analgesic and anti-inflammatory agent comprising as an active ingredient a pharmaceutically acceptable water-soluble salt of incabiraate. 2. The analgesic and anti-inflammatory agent according to claim 1, wherein the water-soluble salt is an inorganic acid salt or an organic acid salt. 3. The analgesic and anti-inflammatory agent according to claim 2, wherein the inorganic acid salt is a hydrochloride. 4. The analgesic and anti-inflammatory agent according to claim 1, which is drug-independent.