JP3064089B2 - New bronchodilator - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a bronchodilator.

[0002]

2. Description of the Related Art 6- (3-Dimethylaminopropionyl) forskolin has the following formula:

[0003]

Embedded image

[0004] The compound is a known compound represented by
-10783). It is not known that this compound has a direct relaxing effect on bronchial smooth muscle. Heretofore, as a bronchodilator, a β-adrenergic receptor stimulant, a xanthine-based drug and an anticholinergic have been used in clinical practice (Japanese clinical practice: 45 (8), 174-180, 1987).

[0005]

However, β-adrenergic receptor stimulants are resistant to drugs by continuous use (J.
Allerg. Clin. Immunol .: 59, 383, 1977), the need for xanthine drugs to monitor blood levels to prevent side effects (asthma: 1 (1), 66, 1988), and anticholinergic drugs Have weak points such as a weak direct relaxing action on bronchial smooth muscle (asthma: 1 (1), 80, 1988), and new bronchodilators are desired.

[0006]

The present invention relates to 6- (3-dimethylaminopropionyl) forskolin (hereinafter referred to as "NK").
H477 ”) as an active ingredient. When NKH477 is used as a bronchodilator, injections, inhalants, tablets, granules, fine granules, powders, capsules, suppositories, alone or mixed with excipients or carriers,
Formulations such as eye drops, patches, ointments and sprays are administered orally or parenterally. Pharmaceutically acceptable excipients and carriers are selected, and their type and composition are determined by the administration route and administration method. For example, in the case of injections, physiological saline, various buffers, glucose
Saccharide solutions such as water, inositol, mannitol and xylitol; and glycols such as ethylene glycol and polyethylene glycol. Further, a lyophilized preparation is prepared together with an excipient such as glucose, inositol, mannitol, xylitol, maltose, sucrose and the like, and an excipient such as phenylalanine. For example, it can be administered after dissolving in a liquid for intravenous administration such as sterilized water, physiological saline, glucose solution, electrolyte solution, amino acid and the like.

The content of NKH477 in the preparation varies depending on the preparation, but is usually 0.1 to 100% by weight. For example, in the case of an injection solution, it is usually preferable to contain NKH477 in an amount of 0.1 to 5% by weight. The dosage is determined according to the age, body weight, symptoms, treatment purpose, etc. of the patient, but the therapeutic amount is generally 0.001 to 30 mg / kg / day for parenteral administration and 0.003 to 3000 mg for oral administration
/ kg / day.

[0008]

The effect of the bronchodilator NKH477 of the present invention is shown in the following experimental examples. Experimental Example 1 The diastolic effect on isolated guinea pig tracheal smooth muscle was assessed by resting tone and the typical tracheal smooth muscle contractors mesacholine, histamine and leukotriene D.
_The relaxation effect on contraction by ₄ was examined using the index. Experimental method After killing Hartley male guinea pigs (Japan SLC), the trachea was removed. The tracheal chain specimen was prepared by carefully forming 20 tracheal pieces and connecting the 5 pieces alternately with a thread so as not to damage the tracheal smooth muscle. Specimens were modified Krebs-Hen
0.5 in a 15 ml organ bath filled with seleit nutrient solution
g The suspension was applied under heavy tension. The nutrient solution is maintained at 37 ° C and 95
% O ₂ + 5% CO ₂ gas was ventilated. The experiment was performed after the equilibrium was maintained for 1 hour. The reaction was recorded isotonicly on a servo recorder (R-12M, R-201) via a transducer (Nihon Kohden, T-111). (1) Relaxing effect on resting tone: NKH477 (10
^-10 to 10 ^-5 M), isoproterenol (10 ^-10 to 10 ^-5
M) and forskolin (10 ^-10 to 10 ^-5 M) are cumulatively administered, and the relaxation response is reduced to 1% by papaverine (10 ^-4 M).
The relaxation rate was expressed as 00%. Moreover, beta _2-adrenergic receptor blocker propranolol (10 ^-6 M) NK in the presence of
The relaxation effect of H477 was also compared with isoproterenol. The concentration (IC ₅₀ ) that relaxes the tracheal smooth muscle by 50% was calculated from the respective concentration-effect curves by the least squares method using the simplex method. Table 1 shows the results.

[0009] (2) methacholine, relaxing effect against histamine and leukotriene D ₄ Shrinkage methacholine (3 × 10
^-6 M), histamine (10 ^-5 M) or leukotriene D ₄
(10 ^-8 M) was administered, after contraction was stabilized NKH477 (10 ^-10
~ 10 ^-5 M) and forskolin (10 ^{-10 -10 -5} M) were cumulatively administered, and the relaxation response was expressed as the relaxation rate, with the relaxation by papaverine (10 ^-4 M) as 100%. The concentration (IC ₅₀ ) that relaxes 50% against contraction by a contracting substance was calculated from each concentration-action curve by the least squares method using the simplex method. Table 2 shows the results.

[0010]

[Table 1] _{──────────────────────────────────── IC 50 (-log [M]} ) NKH477 Isoproterenol Forskolin (number of cases) (number of cases) (number of cases) ───────────────────────────────── ─── Resting tone 8.34 ± 0.21 9.06 ± 0.19 8.05 ± 0.29 (4) (4) (3) Addition of propranolol 8.34 ± 0.11 6.17 ± 0.16 − (4) (3) ─────────── ──────────────────────── Values indicate mean ± standard error

[0011]

[Table 2] _{──────────────────────────────────── IC 50 (-log [M]} ) NKH477 Forskolin (Number of cases) (Number of cases) ──────────────────────────────────── Histamine 7.52 ± 0.14 7.10 ± 0.08 (4) (3) Methacholine 6.33 ± 0.15 6.01 ± 0.08 (4) (3) Leukotriene D ₄ 7.50 ± 0.04 7.19 ± 0.01 (3) (2) ───────────── ─────────────────────── Values indicate mean ± standard error

As is clear from Table 1, NKH477 exerts a powerful tracheal smooth muscle dilating action even in the presence of 10 ^-6 M propranolol, ie, in the state of beta-adrenergic receptor blockade, unlike the beta-adrenergic receptor stimulant isoproterenol. Have. As is clear from Tables 1 and 2, NKH477 has a tracheal smooth muscle dilating action 2-3 times stronger than forskolin.

Experimental Example 2 Bronchodilation by intravenous administration were investigated the inhibitory effects of airway narrowing by leukotriene D ₄ inhalation anesthesia guinea pigs index. Experimental method Male Hartley guinea pigs (Japan SLC) were used. After anesthesia with pentobarbital (40 mg / kg ip), tracheostomy was performed, a tracheal cannula was attached, and artificial respiration was performed with a small animal respirator (Shinano Seisakusho, SN-480-7). Tidal volume is 10 ml / kg, ventilation rate is per minute
60 times. Pulmonary inflation pressure (PIP) is measured as an index of bronchoconstriction by a pressure strain gauge (Nihon Kohden, AP-621G) through a pressure transducer (Nihon Kohden, TP-200T), and a recticorder (Nihon Kohden) , RJG-4124). Inhalation of leukotriene D ₄ is performed using an ultrasonic nebulizer (Nihon Kohden, TUR-3
000) was carried out for 30 seconds from 45 minutes after administration of the anesthetic using a modified inhalation load device for small animals. The concentration of the leukotriene D ₄ solution at that time was 300 ng / ml. Succinylcholine (5 mg / kg, sc) was used to reduce the variation in secondary reactions caused by spontaneous breathing or thromboxane production.
) And indomethacin (10 mg / kg, was administered iv) respectively leukotriene D ₄ inhalation after 20 minutes before and 10 minutes before. The test drug and indomethacin were administered via a polyethylene tube with a needle inserted into the hind limb and forelimb vein. Test drug was injected intravenously over a period of 20 seconds leukotriene D ₄ inhalation start 1 minute ago. A physiological saline solution was used as a control.

Analysis of the Results The ED ₅₀ of the bronchodilator effect at 1.5 minutes after the start of inhalation of leukotriene D ₄ (the dose that suppresses the PIP elevation by 50% in the control group by leukotriene D ₄ ) was determined from the dose-response curve of each group by the simplex method Calculated by the least squares method. Results The results are shown in Table 3.

[0015]

TABLE 3 ──────────────────────────────────── Drug ED ₅₀ specific activity ([mu] g / kg) (n mol / kg) ──────────────────────────────────── NKH477 6.51 11.9 1 Aminophylline 8130 18200 1 / 1500 ────────────────────────────────────

As is apparent from Table 3, NKH477 showed a 1500-fold stronger bronchodilator effect of aminophylline by intravenous administration. Experimental example 3 The bronchodilator effect of inhaled administration was examined with reference to the inhibitory effect of histamine inhalation on airway narrowing in guinea pigs.

Experimental Method Using male Hartley guinea pigs (Japan SLC), experiments were conducted in a constant temperature room at 25 ° C. Under spontaneous anesthesia, each guinea pig first inhales physiological saline into the respiratory tract twice at 15-minute intervals for 30 seconds.
w) was measured 2 minutes after the start of inhalation, and the average of the two measurements was ba
seline SGaw value. Histamine 0.0 after 15 minutes
5, 0.071, 0.1, 0.14, and 0.2% (concentration in the nebulizer, the same applies to the concentration of the test drug indicated below)
Inhalation was performed at 15-minute intervals for 30 seconds until the SGaw value at 2 minutes after the start of inhalation (at the peak of airway stenosis) decreased by 40% or more of the baseline value. Histamine dose response line (log concentration vs.
The histamine concentration that reduces SGaw by 40% is calculated from the SGaw reduction rate (linear regression from two points), and the histamine of that concentration is inhaled 5 times for 30 seconds at 90-minute intervals after 90 minutes.
The SGaw value was measured. Experimental groups were set two groups NKH477 Gun及 beauty control group, the first (initial) and 5 th is a saline pretreated inhalation 2 minutes from the histamine inhalation 3 minutes ago (ba
Similarly, when preparing a seline value and dose-response line, physiological saline was inhaled before treatment), and the test drug NKH4
77 0.1,0.71,5 mg / ml, or were pretreated sequentially inhalation similarly saline. The inhibition rate of airway constriction due to histamine inhalation by the test drug is (initial SGaw reduction rate-SGaw reduction rate under test drug pretreatment) / (initial SGaw reduction rate) x 100
It was calculated as Intact and SGaw values of guinea pigs under spontaneous respiration were measured using constant-volume bodyplethysmo-graph. In other words, individual animals are two-compartment ch
amber and the respiratory flow rate is controlled by a respiratory flow transducer (Nihon Kohden, TV-142T) and a differential pressure transducer (Nihon Kohden, TP-602T).
AR-601G) and a DC amplifier (Nihon Kohden, AD-601G incorporating a 6 Hz high cut filter circuit). In addition, ventilation unit (Nihon Kohden, AQ-601G) and instantaneous heart rate unit (Nihon Kohden,
AT-600G) was used to measure ventilation and respiratory rate. The chamber to which the animal was fixed was placed in a plethysmograph, and an amplifier (Validnyne, MP45) was used to drive the amplifier
dyne, PA701) and plet through a DC amplifier (Nihon Kohden, AD-601G incorporating a 6 Hz high cut filter circuit)
The pressure inside the hysmograph was measured. The change in internal pressure is caused by reducing the sensitivity to 1/10 with individual guinea pigs
Calibration was performed by rapidly placing ml into the plethysmograph and indicating as air volume. Respiratory flow rate, plethysmograph internal pressure, respiratory rate and ventilation volume are recorded on a 4-channel lecticorder (Nihon Kohden, RJG-4124) (for monitoring), and at the same time, respiratory flow rate and internal pressure are digital on the X-axis and Y-axis on the X-axis. Input to Storage Corp. (Iwatsu Electronics, DS-6411) and XY Recorder (Graph-tec
, WX2400), and SGaw (1 / sec · 1 / cm H ₂ O = slope) from the slope connecting the two points of the beginning of the suction phase of the XY loop and one third of the maximum flow velocity with a straight line. (ml / sec /
ml) / (calculated atmospheric pressure -64 cm H ₂ O) SGaw. The atmospheric pressure was read directly from an aneroid barometer (Isuzu Seisakusho). Inhaled administration of histamine and test drug
The aerosolized solution with an ultrasonic nebulizer (OMRON, NE-U11B) (the liquid volume in the nebulizer was 25 and 10 ml, respectively) was sucked by an air pump (Iwaki, AP-032ZN) into a two-compartment chamber (guinea pig). At the tip of the nose) through a silicone tube and an 8-way branch pipe with a cock (for switching the aerosol flow) at a flow rate of 1 l / min. At this time, the air flow rate was adjusted using a desktop type flow meter (Ono Seisakusho).
After inhalation of histamine for 30 seconds, air without aerosol
After sending the remaining histamine aerosol in the tube at 1 l / ml for 20 seconds, the plethysmograph was once opened to the atmosphere to make the atmospheric pressure and the internal pressure the same, and then the plethysmograph was closed to measure SGaw. At times other than measurement, air was constantly sent into the chamber at 1 l / min to prevent guinea pig oxygen depletion.

Analysis of Results EC _{50 of} bronchodilation (airway stenosis caused by histamine was reduced by ₅₀ %)
% Inhibitory concentration) is a dose-response curve using the mean
It was determined using the method of least squares by the line or we simplex method. In the control group, no suppression of 10% or more was observed on average. Results The results are shown in Table 4.

[0019]

[Table 4] 薬 物 Drug EC ₅₀ (mg / ml) (× 10 ^-3 M) ──────────────────────────────────── NKH477 0.714 1.31 ────── ──────────────────────────────

As is clear from Table 4, NKH477 exhibited a bronchodilator effect even by inhalation administration. Incidentally, LD ₅₀ values of NKH477 for male rats was 8.9 m g / kg intravenously.

[Effect] As is clear from the above, NKH477 has a potent bronchodilator effect that is effective even in the β-adrenergic receptor-blocked state, and various bronchodilators, that is, various symptoms based on airway obstructive disorders of various diseases Is expected as a drug that relieves the disease.

[0021]

【Example】

Formulation Example 1 Purified water is added to 30 parts by weight of NKH477 to make the total amount 2000 parts, which is dissolved, and then sterilized and filtered using a Millipore Filter-GS type. 2 g of the filtrate was placed in a 10 ml vial and freeze-dried to obtain a freeze-dried injection containing 30 mg of NKH477 in one vial.

Claims (1)

(57) [Claims] (1) 6- (3-dimethylaminopropionyl)
Bronchodilators containing forskolin as an active ingredient (excluding antiasthmatics).