TWI228988B - A vasorelaxant alpha/beta-adrenoceptor blorker with distinguish antioxidane - Google Patents

Abstract

The invention disclosed some derivative chemically with Vanilloid based moiety. As formula I and II wherein R1 is selected from -C3-6COC1-5, unsaturated C3-6 alkyl chain, or -C1-5COOC1-5. R2 is selected from C3-6, -C1-6O(C6H4)-2-OC1-3, -C1-6O(C6H4)-3-OC1-3, -C1-6O(C6H4)-4-OC1-3. Through in vivo experiment to prove those compounds have generation alpha/beta-adrenoceptor antagonist with vasorelaxant activity by inhibiting Ca2+ channel, receptor-mediated Ca2+ mobilization and by K+ channel opening, and to have additional potentially antioxidant effects.

Description

1228988 ------ 、 Explanation of invention (1) Field of industrial application The present invention discloses a series of Vanilloid derivatives, which have ▼ type and type B adrenal cortex receptors (α / $ Touch⑼㈤mail⑽) binding affinity, antioxidant activity. Jing Bei reported in 1999 by Huang YC. Et al. In J Cardiovasc Pharmacology Vol. 34, pages 10-20, and 1999 by Huang YC. Et al. In Drug Development Research, Vol. 47, pages 77-89. Eugenodilol, ferulidil〇1, vanidipinedilol, flavor] iabecjipinedilOL-A and carvedilol are emerging third Substitute type B blocker ($ _biocker), has better vasorelaxant and antioxidant activity than other type B blockers. These new discoveries have inspired us to further design new compounds with potent antioxidants and α / B adrenoceptor blocking activity. Page 5 1228988 V. Description of the invention (2) During the past ten years, the inventors have developed a variety of type B blockers with the basic structure of guaiacol valerinol (Vanil_ (heart bl. cker). These blockers are derived from natural products, for example, Chen IJ et al., 1993, Gen Pharmacd, Vol. 24, pp. 1425-1433, reveal new compounds derived from xanthone 'and 1994 Chen IJ, et al., In Journal of Medicinal Chemistry, Vol. 37, pp. 938-943, revealed a series of new compounds derived from capsaicin. Sheu mm, et al., Pharmacology, Vol. 54, p. 211-224 Page reveals vanillin derivatives, dehydrozingerone, and bamboo biology unveiled by Wu BN et al., Asia Pacific Journal of Pharmacology, Vol. 11, pp. 5-12, 1996 1996 Chen SJ et al. Revealed new compounds derived from eugenol in Drug Development Research, Vol. 40, pp. 239-250. Chen IJ, et al., British Journal of Pharmacology, Vol. 119, pp. 7-14 Arylhydroxypropanolamine (guaiacoxypropanolamine), a guaiacol type B blocker, these new compounds all have 3-methoxy-4_hydroxoxy (3-methoxy) -4-hydroxybenzyl) group.

Page 6 1228988 V. Description of the invention (3) Object of the invention The primary purpose of the present invention is to reveal a series of newly synthesized novel vanilloid derivatives which have A / B adrenocortical receptors & / Adrenoceptors) binding affinity (bindingafgnity), antioxidant activity. Another object of the present invention is to disclose a series of conventional compounds, which have been confirmed by animal experiments to have A / B adrenocortical receptors (α // adrenocceptors) binding affinity and antioxidant activity. Those skilled in the art will undoubtedly know the objects and advantages disclosed by the present invention after reading the following detailed description of the preferred embodiments shown by different diagrams. i Detailed description of the invention The present invention discloses a series of Vanilloid derivatives, which have binding affinity of A and B adrenoceptors, Oxidation activity. Such as Formula I, Formula π

Page 7 1228988 V. Description of the invention (4) Vanilloid series derivatives, the heart of which can be _〇3_6αχν5, a linear C3_6 containing unsaturated groups, and -CwCOOCw, R2 2-OCV3, -CmOCQ ^ M -OCm, -〇ν60 ((36Η4) -4- (χν3. Using compound 1-6 as raw material, reacting with epichlorohydrin in an alkaline environment to obtain propanolamine, and then separately with isopropylamine (isopropyl amine), tert-butylamine or guaiacoxyethylamine are reacted to obtain compounds 7-16 having a Vanilloid structure as shown in Table 1. The above compounds 1-6 are For example, the derivative of the formula I, R1 may be -c36COCi 5, a linear C% containing an unsaturated group, and CwCOOC ^; and the substituents listed in Table 2 are specific derivatives among them.

OH

Formula Π Formula I The synthesized product was subjected to elemental analysis, mass spectrometry (MS), infrared spectroscopy (IR), hydrogen nuclear magnetic resonance spectroscopy (iH-NMR, CDC13),

1228988 '"X; State (5) ^ "' ----- Physical and chemical information such as ultraviolet absorption (uv). The effect of pharmacological activity is evaluated by appropriate experimental models to determine the activity of the compound. The compound of the present invention is added with various excipients, carriers, or diluents as required, and a pharmaceutically acceptable acid can be added to form a test composition. These preparations may be solid, liquid, or parenteral injection forms for oral administration, rectal administration, or ointment forms directly applied to the affected area. The solid age method is known to be made into tablets or filled with capsules by adding dispersing agents such as Tianyu powder, Linmethylcellulose, or binders such as ethanol and glycerin, or stearic acid and lactose. Or made into a fine body preparation of suppositories. The aqueous solution and the salt solution of the compound of the present invention are used to adjust the acidity of the linate-based buffer solution to make the pH value reach a proper level, and the additives or emulsifiers are added to make injections or other additives. The hair compound or the permissible acid addition salt can be mixed with various bases according to a conventional preparation method and can also be made into an ointment dosage form. Using the compound of the present invention as the domain money to prepare the school ridges, mammals and other mammals produce the main effect of the main ingredients. Generally, it can be formulated according to the needs of symptoms, usually 50 to 300 per person per time. 3 mg daily-owed. 1228988 V. Description of the invention (6) An isomer of eugenol, whose structure is {4- [2-hydroxy-3- (2-methoxy-1-oxyethyl gas basically) propoxy]- 3-methoxypropanylbenzene ({4- [2_11 > also (^ -3- (2-methoxy-1 -oxyethylamine obenzene) propoxy] -3-methoxy}-1-propylenyl-benzene) is iso clove oil Phenoldiol (isoeugenodi10i). A 3-methoxy-4-hydroxyoxy group (3_niethoxy-4-hydroxy) group with the basic structure of guaiacene, in 1993 by Raj akumar DV et al. In Biochemical Pharmacology Volume 46, pages 2067 to 2072 reveal that isoeugenol can inhibit the effects of arachidonic acid-induced lipid peroxidation (iipidperoxidation) and antiaggregation (isoaggregation). Synthesized from iso-syringene oil (isOeUgenOl), chemically includes aryloxypropan in the basic structure of adrenoceptors_blockers. l), a guaiacolethylamine group with the basic structure of a type A blocker. The inventors discovered that it is derived from isoeugenol FIG alcohol of iso eugenol bis (isoeUgen〇l〇l) shown in FIG. 1, belonging to the same manner as beta blockers (/ 3 -blocker). However, in 1999 Lin YT et al Jpn J Pharmacol 80

Page 10 1228988 V. Description of the invention (7) Pages 127 to 136 reveal that the compound has smooth muscle relaxant activity in the trachea and vasculature and no A-blocker activity. From experimental observations, patients with heart failure under considerable oxidative stress, and under this pressure, show an increase in the composition of oxygen free radicals, which can lead to the death of myocardial cells or apoptosis. It is more effective than other beta-drugs. The derivatives of formula I and formula II prepared according to the method of the present invention also have the same characteristics as the antioxidant, like the derivatives of Vanilloid series, isoeUgeno scent (15). Activity, and attenuates possible apoptosis. According to the present invention, the derivatives of formula I and formula π have the following pharmacological activity characteristics, including antihypertensive effect, binding affinity between type 曱 and type B adrenal receptors (α / cold adrenoceptors). , Essential sympathomimetic activity (isA, Intrinsic Sympathomimetic Activity), vascuiar smooth muscle relaxant activity (vascuiar smooth muscle relaxant activities), and especially possesses antioxidant (antioxidant) activity.

Page 11 1228988 V. Description of the invention (8) The rat's heartbeat in vivo is described in detail. The normal blood pressure of male rats (Normotensive Wistar strains (NTRs) weighing 200 ~ 300 grams) was selected through pentobabarbital sodium. ) Intraperitoneal anesthesia, perform a tracheotomy and insert a catheter to keep breathing smooth. In the left vein, femoral veins were inserted with polyethylene catheters to "administer Tetra Pak, and a 3-way stopcock was used, and one end was connected to the device to inject the drug into the syringe barrel. ; The other end of the connecting device is a syringe of physiological saline solution, and a small amount of physiological saline solution can be injected immediately after administration to prevent the drug from staying in the polyethylene tube and affecting the accuracy of the experiment. The tube is also a three-way biopsy. One end is connected to a heparin solution. When a plug is found in the polyethylene tube, it can be used to clear the tubing. The other end is connected to a pressure transducer (Model P10EZ; Spectramed, Oxnard, CA, USA), and then record the systemic arterial pressure, mean arterial pressure, and heart rate with a recorder, in this way, the blood pressure and heartbeat effect of the drug can be evaluated in rats. The rats are administered 0.1, 0.5, 1.0, 3.0 mg / kg and other doses of isoeugenol

Description of the invention on page 12 After (9) (isoeugenodilol, 15) (001), the differences in heartbeat and blood pressure were compared. Rats with normal blood pressure administered different doses of the compound isoeugenol bisol (isoeugenol Isoeugenodiiol, 15), as shown in Figure 2, can cause a slow heartbeat and continuous blood pressure reduction for up to about 1 hour. This heartbeat slowing and blood pressure lowering effect is positively correlated with dose. In a ganglion-blocked anesthetized rats experiment, 0.5 mg / kg of isoeugenodilol and 1.0 mg / kg of propranolol were administered intravenously. , Can reduce 0.5 pg / kg (-) L · isoproterenol (isoproterenol) -induced accelerated cardiac response, from 105.6 to 5.8 bpm beats per minute (beats mine-l) significantly reduced to 7L3 ± 6.7 bpm, or Significantly reduced from 103.3 to 8.8 bpm to 36.7 ± 9.9 to 0111 (^ < 0.01511 = 8). The rats were first administered to the snake root test (1 ^ 611) 1116), and then intravenously injected with 10 gg / kg of phenylephrine and L-phenylephrine could raise hypertension by 57.5 ± 4.5 mmHg. If administered with 0.5 mg / kg Isoeugenol bisphenolate 〇61 ^ 11〇 (1 丨 1〇1,15) can be found to significantly reduce the drop response to 31.9 soil 10.6 11111111 eup (ρ < 0 · 015 n = 8), but it is not affected by 1.0 mg / kg prote

Page 13 1228988 V. Description of the invention (10) (propranolol) The blood pressure is still 57.0 ± 4.9 mmHg.加速 The accelerated cardiac response induced by L-isoproterenol, and the hypertensive response of phenylephrine (L-phenylephrine) are inhibited by the influence of isoeugenodilol (15), It is clear that isoeugenol (15) has a blocking effect.

Antihypertensive effect Spontaneously hypertensive rats (SHRs) weighing 200-300 g in males are indirectly anesthetized with a tail cuff method using a rat tail pressure gauge and a flow meter (man 〇meter — tachometer) (NATSUME KN-210, Natsume Seisakusho Co. Ltd.

Tokyo, Japan). Measurement of systolic blood pressure and heart rate. Place mice in a Plexiglass holder maintained at 37 ° C for 15-20 minutes to increase body temperature, expand the caudal artery, and pressurize to 4 ° degrees to detect the beat.

1228988 V. Description of the invention (11) For the average value, only rats whose systolic blood pressure is not less than (^) 180 mmHg are used. Oral month 10 mg / kg, 30 mg / kg, and 100 mg / kg before or after a single dose of isoeugenodilol (isoeugenodilol, 15), respectively, at 0.5 hours, 1 hour, 2 hours, 3 hours, 4 Hours, 5 hours, 6 hours, and 24 hours were measured. • The unadministered control group was replaced with physiological saline.

Results Spontaneous high-pressure A rats without anesthesia (conscious) received oral doses of isoeugenodilol (15, isoeugenodilol) as shown in Fig. 3. Both the blood pressure drop and the heart rate slowdown showed dose dependence. Hypotension and bradycardia appear 12-24 hours after using vanidipinedilol.

Evaluation of Type A and Type B Adrenal Cortex Receptor Blockers (α-/ / 3 -adrenoceptors-blockers). Normally normotensive male rats (NTRs) are administered intravenously with ganglion-blocker 5 mecamylamine. Make sure it matches the initial heartbeat. By thigh

Page 15 1228988 V. Description of the invention (12) A single dose of isoeugenodilol (15) is administered intravenously. Ten minutes later, L-isoproterenol was injected again and the response in the control group was expressed. In addition, mice were pre-administrated with 5 mg / kg reserpine intravenously, 10 pg / kg phenylephrine was administered intravenously 24 hours ago, and then 0.5 mg / kg isoeugenol was administered intravenously 15 minutes later (15) or 10 mg / kg propranolol. After 10 minutes, phenylephrine (rineL_phenylephrine) was injected again. Evaluation of the relaxation effect of life pipes: Take 300-450 grams of guinea pigs, knock them from the head with force, bleed them from the carotid artery, quickly remove the thoracic artery, and keep it cold. In the solution, carefully remove the fatty connective tissue around the blood vessel wall, and then cut the thoracic aorta to a length of about 5 mm. The aortic ring was fixed up and down with two “H” -shaped platinum wires, and placed in a millilitre tube. In tissue tanks with a mixture of 95% oxygen and 5% carbon dioxide, one end is fixed to the Wei Wei of the organization. The force transducer ii (f_ transducer) is used to record the isometric contraction via a recorder (is〇metric c〇ntracti〇n ). The sample was given 10 # Μ phenylephrine (㈠ grams of tension, 60 minutes after equilibration, first 1228988

phenyiephrine) Secret of working age, wait for it _ wash off the phenylephrine and give it to M phenylephrine to achieve maximum contraction, after it is balanced, give 10.6 or 10 ~ different concentrations of test compounds Observe the inhibitory effect caused by the test compounds at different concentrations.

Color blood vessel relaxation activity ^^ Heart activity, Xi Pinglan set the rat thoracic artery ring in the tissue trough, and add 3 × 10 ～ 6 M norepinephrine or 75 inM potassium chloride (KC1) to Induce

Stable shrinkage. When the reaction reaches its maximum tension, i.e., close to 1QQ%, isoeugenodilol (15) is added to produce relaxation. 75Fu κα solution can also be modulated by 70 mM NaCl instead of isoimoiar κα, in an attempt to clarify the possible mechanism of isoeugenol bis (15) relaxation. Isoeugenodilol (15) was used to evaluate the relaxation of the following compounds; 〇 "μΜ-a voltage-dependent calcium channel activator Bay K 8644, 10 mM-a non-specific Anon specific inhibitor of K + channel, TEA (tetraethylammonium chloride), 1 μM-a species susceptible to adenosine triphosphate (Ατρ)

Page 17 1228988 V. Description of the invention (14) adenosine triphosphate) inhibitor glibenclamide, 3 × 10-4M, a nitric oxide synthase inhibitor nitrguanyl vinegar (N ^ nitro-L-arginine methyl ester (L_NAME), and 3 × 10.5 μ

Inhibitor of a staglandin biosynthetic enzyme, indomethacin. For example, in 1997, Chen SJ. Et al., Drug Dev Res., Vol. 40, pp. 239-250, described that isoeugenol (15) is susceptible to norepinephrine in the cell regarding Ca + 2 release and injection ( norepinephrine) sensitivity. Isoeugenol (15) was incubated for 10 minutes in a physiological solution containing 1 mM ethylene glycol-diamine ethyl ether (EGTA), and norepinephrine was added ° Results

Voltage-dependent calcium channel blocking activity in aorta (M) ltage-Denendent Ca + 2 Channel Rlocking Activity on UnMed

Aorta) Aortic contraction induced by 75 mM K + is shown in Figure 4, regardless of whether 10-6 M, 5x10 M, or 105 M iso-eugenodilol (15).

! 228988 month description (15) '· --------- Relaxation effects are dependent on the dose (dose_dependence). 10 minutes before adding ιοΜ Μ Bay Κ 8644 to the tissue slot, adding chlorin unloading, when the shrinkage reached stable, add 10-6 Μ, 5x10.6 Μ, and 10-5μ iso clove oil, respectively. Diols (I noodles 0, 15). As shown in the figure), Bay κ Li is sufficient to affect the κ + induced contraction of iso-syringrolidol (I5) at high concentrations. Iso cloves .. / Dominated by IQ by Pandiol (I5). The value is i + 5 M, and Bay κ 8644 is pretreated: The contraction effect induced by 75 mM potassium chloride is 5 66 × 10-5 M (n = 8). Display · Iso clove oil (15) has the ability to block aortic-dependent voltage Nazi channel activity 0 has a potassium channel opened in the aorta ^^ haimeLQpming ^ iciiyitvon Τς〇ικη Anrfp)

The use of two potassium channel inhibitors, tetraethylammonium chloride (TEA) and glibenclamide, can explore the role of K + permeability. Γ can relax in isoeugenodilol 3χ10.6 Norepinephrine-induced thoracic aortic rings contraction process. As shown in Figure 4B), whether 10-7 M, 5x10-7 M, or 10-6 M isobutyl

Page 19, 1228988 V. Explanation of the invention (16) The isoeugenodilol (15) is sufficient to relax the norepinephrine-induced pre-aortic contraction. This phenomenon is susceptible to 10 mM tetraethylammonium vaporization (TEA). reduce. However, comparing the control group, it was found that 10-6 glibenclamide was only moderately diminished with isosyringrolol (15) to induce relaxation. The ic5G value of isoeugenol diol (is〇eugenodil〇U5) is 4.01X10 · 7

Μ ′ Tetranitride (TEA) is 7.08 X 10 Μ, glibenclamide

The ic% value of inhibition of norepinephrine pretreatment-induced contraction was 8.61 X 10-7 M (n = 8). In contrast, the following experiments were performed using nitrate (L-NAME) and indomethacin to determine whether nitric oxide (NO) and prostaglandin I2 (PGy were involved. However, exposure to 3x10.4 M l-nitroguanidyl ester and 3 × 10_5 M indomethacin will not be meaningfully reduced. The 0.6M isoeugenodilol 515 inhibits norepinephrine (nor sentence> inephrine) pretreatment. Induced contraction. Norepinephrine (vomiting in India hrine) induces _ purpose contraction IM1 page 20 1228988 V. Description of the invention (17) Bipinephrine produces biphasic contractile response, including rapid phase (phasic) And tonic, etc. This experiment is to determine the role of Ca + 2 in isoeugenodilO5 inhibition of norepinephrine-induced contractile activity in isoeugenodilO5. Norepinephrine-induced contraction, whether pretreated with 1CT6M, 5x10-6M, and 10.5M isoeugenol (15), will become smaller in both cases as shown in Figure 5; the first is Ethylene glycol _ di-(/ 5-aminoethyl scale) N, N'_ tetraacetic acid 111} such as 1 ^ 017〇) 1-1 ^ (/ 3 · amineoethyl Ether) N, N'-tetraacetic acid). Calcium-free physiological salt solution of norepinephrine (hrepinephrine) produces rapid contraction. The other was the introduction of chlorinated # 5 to produce sustained contraction. Equimolar concentrations of isoeugenol bis (15) inhibit the first phase more strongly and strongly than the second phase. A-Adrenoceptor Antagonism (a-Adrenoceptor Antagonism, cf. Huang YC. Et al. 1999 J Cardiovasc Pharmacol. Vol. 34 pp. 10-20) or 1999 Drug Devel Res vol. 47 p. 77

Page 21, 1228988 V. Description of the invention (18) to 89. The report describes the evaluation of the mechanism of action of isoceugenodilol (15) on a radrenergic and pigment-activated (sertonjjjergic). Method 'A Wistar rat's thoracic aorta rings were used. The aorta was suspended in a physiological saline solution at 37 C at a resting tension of less than 1 g. The physiological saline solution: Composition of sodium gasification (118 mM), potassium chloride (4.8 mM), gasification words (2.5 mM), magnesium sulfate (1.2 mM), potassium squamate (1.2 mM), sodium bicarbonate (24 mM), glucose (glucose, 11 mM). In the "adrenergic blocking action" test, after 30 minutes of administration of isoeugenol bis (15) solution, 10.1 () to 10.4 were added gradually. Norepinephrine was used to induce contraction. In the course of two consecutive administrations of norepinephrine, it was washed three times with saline and then re-equilibrated for i hours, maintaining the physiology after an interval of 10 minutes each time. Freshness of saline solution. Calculate the percentage of maximum control response as the response of norepinephrine, and observe isoeugenol

1228988 V. Description of the invention (19 ^ ~ 1 ~ ---------------- Blocking the pigment preparation activity, after giving 5 mM isosyringa oleyl bishydrin ⑽ solution 30 knives, gradually add 1 0-7 to 10-4 M soluble serotonin to induce contraction. Result

As shown in the figure, the relaxation effect of isoeugenodilol, whether 10-8 M, 10-7 M, or 10-6 M, exhibits a concentration-dependence and concentration-dependent effect. Competitively inhibits norepinephrine-induced contraction. Isoeugenodilol (15) was dose-dependent and similarly caused norepinephrine concentration-response curves to shift to the right. Of isoeugenol bis (15)? The value of 8-2 is 7.47 ± 0.45, and the labetalol at the mouth of the arterial ring is value of 8.87 ± 0.08. Table 3 is the pA2 value and slope regression line of isoeugenol (15) and laxta. However, serotonin has no effect on the concentration response curve of isofluorol (15) at 10 · 5M as shown in the figure) ^ ___

Page 23 1228988 V. Description of the invention (20) Antibodies against β-1 and β-2 adrenal receptors (βr and β ^ -Adrenoceptor Antagonism) cf. Huang YC. Et al. 1999 J Cardiovasc Pharmacol.

Vol. 34, pp. 10-20, or 1999 Drug Devel Res., Vol. 47, pp. 77-89, and the method described by Yeh JL. Et al. In 1999 at J Cardiovasc Pharmacol. The left atrium was erected in a 10-ml organ bath, one end of which was fixed, and the other end was connected to a force transducer (Model FT03; Grass, Quincy, MA? USA ·). The heartbeat rate was measured by the channel i of the frequency-converting amplifier (Queequy-converter ampiifier) (Model 13_6615_60; Gould, Valley View, OH, U.S.A.). Two platinum electrodes were placed at two intervals in the left atrium, and the atrium was placed in a Krebs solution at 37 ° C for experiments. The composition of the heart coffee solution is: vaporized sodium chloride (3 mM), potassium vaporized (KC1, 4.8 mM), calcium vaporized (CaCl2, 2 2 _), potassium phosphate (KH2P04, U mM), gas Magnesium (Mgcy 2 Fu), sodium carbonate (NaHc〇,

Page 24 1228988 V. Description of the invention (21) The mixed gas of carbon oxide was extended first to the right atrium and left atrium to reach a basic tension of 0.02 to 0.05 g 'and maintained at equilibrium for 60 minutes. It is estimated to block A adrenergic blocking activity (y5r adrenergic blocking activity), control the concentration-response curves of ㈠L-isoproteren to increase inotropic and chronotropic Effect. Allow the atrium to maintain balance for another 60 minutes. Before the administration of 1 (T1G to 3 × 10 · 6 ㈠ ㈠L-isopr〇teren 〇) gradually increasing concentration (cumulative concentration), the agent was administered at different concentrations. Cultivate. Suspend the trachea of Guinea pig in a 20 ml Krebs solution, pass it into an organ trough with a mixture of 95% oxygen and 5% carbon dioxide, and keep it at 37 ° C. Stable spontaneous tone is achieved. The degree was initially maintained at a tension of 1.5 g for 60 minutes. After contraction at 3 × 10_6 M carbachol, the concentration was about 3 · times ㈠ L-isoproterenol to obtain an increasing concentration · relaxation response Curve (cumulative concentration-relaxant Π !! Page 25 1228988

V. Description of the invention (22) _e) 'Incubate for 3G minutes. The effect of the / 3 radrenoceptor antagonist on the type 2 renal axe receptor antagonist can be measured by a television image. ㈠L · Esopourol inhibits the relaxation of the trachea. The percentage of the maximum relaxation effect is the expression of whether the trachea shows relaxation. result

— ~~ See the adrenal cortex receptor antibody activity (Effects on / 9 iAdrenoceptor Activity, as shown in Figure 7 (\) 'isoeugenocjiioi (i5) antibody to the right atrium of Wistar rats Induced by (-) L-Aesoporo (iSOpr〇teren〇i), it exhibits a time-varying effect. In addition, whether it is 10.8 M, 10.7] y [, or 10_6 M iso clove

Olefinol (15) can shift the concentration response curve to the right to show a concentration-dependence. The ρΑ2 value of iso clove oil in the right atrium is 15 · 33 ± 0.12, the ρα2 value of labetalol is 7.91 ± 09, and the ρα2 value of atenolol is 7.29 ± 0.13, the ρΑ2 value of prOpran〇i〇i is 8.56 ± 0.11. As shown in Figure 78), the left atrial isoeugenol (15) body was stimulated with electricity

Page 26 1228988 V. Explanation of the invention (23) " * ------ will also counteract the positive contractile force effect induced by ㈠L · Esopouro (150 ping 0, such as 〇1), and ㈠L-Esso Popro (isoproteren〇1) produces a dose-dependent (dependent) shift to the right to gradually increase the concentration-response curves. The obvious PA? Value of isoeugenol diol (15) in the right atrium is 7.80 ± 0.09, the pa2 value of labetabl is 7.83 ± 0.07, and the a value of Yaterno is 7. 18 ± 0.05, the pa2 value of Prozac is 8.32 ± 0.09 as shown in Table 3. 'Using the regression line slope value (siopes pi〇ts) to calculate isoeugenol diol (15). Now, the mouth and value of Projbar. Type 2 adrenal cortex sensing_Effects on β ^ -Adrenoceptor Activity

As shown in FIG. 7 (C), the spontaneous tone of the trachea of a guinea pig (Guinea pig) treated with blood pressure level, regardless of the concentration of 10-8M, 10 · 7M, or 106M, is expected. Diols (isoeUgeno (jil〇l5l5) can all compete against (-) L-isoproterenol) induced relaxation. Isoeugenol (15) similarly makes concentration-response curves ) Produces a rightward panning phenomenon. Isoeugenol (15) is more than atenolol

Page 27 1228988 V. Description of the invention (24) It is more potent and significantly less potent than propranolol, like a type 5 adrenal cortex receptor blocker (/ 5 radrenoceptor blocker). The obvious isoeugenol diol (15) in the trachea has a pA2 value of 7.26 ± 0.11, a ρΑ2 value of labetalol of 7.54 ± 0.16, and a ρΑ2 value of atenolol of 5.85 ± 0.12 The PA value of Procamba is 8.27 ± 〇12. Isoeugenol diol (iSeugenodilOU5) selectivity ratio of ethylene type / ethylene type (; 3m / β2) is 12, the selectivity ratio of derivative 10 is .0, the selectivity ratio of derivative η Is u, implying that all three derivatives are a non-selective type B adrenocortical receptor blocker. As shown in Table 3, there is no significant 1.0 difference in the regression line slope values (sloppes plots) of the PA2 value and the slope. Calculated from the inverse logarithm of the difference in PA2 values between the right atrium and the trachea, the selectivity ratio of ethylene / ethylene type II, the ratio of isosyringadiol is 12 " the ratio of the female tower is 2.3 ', obviously the right atrium is more effective than the trachea Force, and both are effective forces, so as shown in Table 4, the order of selectivity of type B- / B-II is Yatino (labetalol) > 普罗 te (pr〇pran〇1 (1) > Isoeugenodilol (15).

Page 28 1228988 V. Description of the invention (25) Essential sympathomimetic activity (isAJntrinsic Sympathomimetic

Activity) 24 hours before the experiment, according to the data of Yeh JL. Et al. In 1999, the animals were pretreated with 10 mg / kg reserpine intraperitoneally, and the right atrium and left atrium of rats and trachea of guinea pigs were removed. Obtained with L-isoproterenol, propranolol, or isoeugenodilol (isoeugenodilol, 15), concentration-response curve Cumulative addition. Results The lack of essential sympathomimetic activity is shown in Figure 8. Iso clove oil is expected to be diol (^ 1 ^ 11〇 (1 丨 101515), propranolol, and L-Isopolna ( isoproterenol) dose-response curves of the right atrium frequency and left heart * tension. L-Aesoporol produces a maximum concentration-dependent increase of 3 × 10 · 6 Μ in heartbeat and contractility. Isoeugenol (15) and prolobar do not increase heartbeat, do not induce negative contractility and time-varying effects. Usually these two compounds increase

Page 29 1228988 V. Description of the invention (26)-The concentration process suddenly interrupts the dependence effect, resulting in the figure shown in the figure) and (printed, preventing the organs from stimulating or not stimulating at 10] V [and 10 Μ / 辰). On the other hand, as shown in Fig. 8t), the concentration of iso-syringarepanol (15) in the concentration of 10-4 M has almost no essential relaxation in the trachea.

Adrenoceptor-R 彳 r ^ jng Assay Rats (Wistar rats (250_400 g)) were sacrificed and decapitated, according to Huang YC et al., J Cardiovasc Pharmacol, Vol. 34, Vol. Pages 0 to 20, described in Drug Devel Res, Volume 47, Pages 77 to 89 in 1999, quickly separated ventricle, lung and brain tissue. Protein content was determined according to the method described by Bradford's in 1976 '. Carry specific stone

Combination of adrenal pimple susceptor to / 5-adrenoceptor antagonist [3H] CGP-12177. [3H] CGP-12177, ventricular membranes or lung membranes (200-300pg) in 75 mM Tris-HCl buffer solution at 25 ° C 60 minute. Tris-methylaminomethane hydrochloride (Tris_HCl) buffer system includes 25 mM magnesium chloride with or without 10 μM propranolol to a total volume of 250 μΐ.

Page 30 1228988 V. Description of the invention (27) Competitive binding experiment; Add the competitor directly to the culture medium, add 1 ml 〇f ice-cold analysis buffer, and measure the culture medium, and immediately use a glass fiber fishing vessel (Whatman GF / C glass fiber filters) ^ # ^^ t (port filter manifold) (Millipore, Bedford, MA, USA). The filter was immediately washed three times with 5 ml of ice-cold buffer, and 4 ml was added before drying in an oven at 80 ° C for 2 hr.

Triton-toluene based scintillation fluid. Membrane_ Combined with [3H] CGP-12177 set in the filter, counted by Beckman LS6500 trace liquid system (Fullerton, CA, U.S.A.), to achieve 45% efficiency. In each experiment, membrane protein and [3H] CGP-12177 were cultured at 10 μM propranolol, and binding of non-specific [3H] CGP-12177 was determined. The [3H] CGP12177 binding total of each test substance minus the non-specific binding number is the specific binding value 0

Specific 曱 -type adrenal cortex receptors (α adrenoceptor antagonist) in combination with [3H] prazosin, compared with the cold-adrenal cortex receptor assay method. [3H] prazosin was incubated with 250-300 g of meninges in 3 mM trismethylaminomethane hydrochloride (Tris-HCl) buffer for 60 minutes. Trihydroxymethylaminomethane hydrochloride buffer solution includes 10 mM magnesium carbide, with or without addition

Page 31 1228988 Description of the invention (28) Phenolamine to a total volume of 500 μ1. Without specific [3 特定] ρΓ32〇 ^ binding, the 臈 protein and [3fflprazassoin were cultured at 0.1 mM fentonium. The total number of [3H] Pmz〇Sin bindings of each test substance is subtracted from the non-specific binding number, which is the specific binding value. 0 Result The effect of adrenal cortical receptor and radioactive agent binding (Effect on Radioligand Rinding Ass ^ y) is shown in Table 5. The results showed that the binding affinity of isosyringadiol (isoeUgen〇cjii〇i, i5) -log 1C% value, and the performance of various adrenocortical receptors on the major receptors of type 1 and type 2 adrenal cortex pKi value. In ventricular membranes, isoeugenodilol has a pKi value of 7.36, labetalol's pKi value of 8.39, atenolol's pKi value of 6.58, and Proteolol has a pKi value of 9.64. The expression of isosyringarepan (15) in lung membranes of rats was 7.27, the pKi of labetalol was 7.83, the pKi of Yatenor was 5.07, and The pKi value is 9.26.

Page 32, 1228988 V. Description of the invention (29) Atenolol shows high affinity for Type 1 adrenal cortex receptors, and shows 32.4 times more selectivity for Type 1 / Type 2 receptors; in contrast, isosyringrolol ( 15, isoeugenodilol), labetalol, and propranolol showed lower selectivity for type I receptors, and showed selectivity for type 1.2 / 3.6 and 2.4 times for the type B / type 2 receptors, respectively. . The binding effect of beta-1 and beta-1 adrenoceptor antagonists on [3H] CGP-12177 is in the order of Provencal> Lazarta > isoeugenol (15) > atenolol ). IsoeugenodilO5 and labetalol bind to [3H] prazosin to produce a concentration-dependent inhibition phenomenon. In the rat brain membrane, isosyring oil is mixed. The pKi value of the alcohol (15) was 7.41, and the pKi value of the Lahu tower was 7.28. The order of potency of adrenoceptor antagonists for binding of [B] adrenoceptor antagonists to [3H] prazosin is isosyringrolol (15) > σ spinet tower (labetalol). 2. Measurement of Lipid Peroxidation in Rat Brain Uniformity

Page 33 1228988 V. Description of the invention (30)

Peroxidation in Rat Brain Homogenate) 10 mg rat brain homogenate was placed in a saline solution containing 0.90 / 0 saline to form thiobarbituric acid-reactive substance, TBARS) to determine rat membrane lipid peroxidation. 1 ml rat brain homogenate was incubated with 10 μΐ of compound at 37 ° C for 5 minutes. Following the method described in the report by Huang YC. Et al. In Drug Devel Res., Vol. 47, pp. 77-89, 0.1 ml of 0.25 was added. mM iron chloride (FeCl2), 1 mM ascorbic acid for lipid peroxidation. After 30 minutes of incubation, the reaction was stopped by adding 0.1 ml of 0.2% butyl via toluene (BHT, Butylated Hydroxytoluene). Add thiobarbituric acid, heat in a boiling water bath for 30 minutes, and extract s Barbituric acid-reactant (TBARS) with n-butanol to measure at 532 nm. The thiobarbituric acid-reactant total was quantified by linear regression with malondialdehyde standardization. Data Analysis and Statistical Evaluation experiments, with or without the addition of agonist median effective concentration (EC5Q) value to obtain the dose ratio i ^ an ^ s page 34 1228988 V. Description of invention (31) rate. According to the calculation method of Furchogott [1972], the antibody constants of Type B and Type II adrenal cortex receptors (Dissociation constants, KB) KB = [antagonist] / (dose ratio-1) pA2 is the negative logarithm of KB, according to In 1959, Amnlakshana 0 and Schild HO. In Br. J. Pharmacol., Vol. 14, pp. 48-57 reported on the method of calculating slope regression lines. According to the radiation-binding analysis, Kd stands for the dissociation constant of radioligand, Bmax stands for the maximum binding capacity, and Ki stands for inhibition constant. 1949 Scatchard G. in Ann. NY Acad. Sci. Vol. 51 pp. 660-1011 reported that non-linear curves using the Scatchard analysis method were determined using the LIGAND program. The Kd value of [3H] CGP_12177 combined with Type I adrenal cortex receptors was 0.16 ± 0.03, and the Kd value of combined with Type II adrenal cortex receptors was 1.21 ± 0.15 nM. At 25t: [3H] CGP-12177 combined with type B adrenal cortex receptor Bmax is 43.1 ± 2.5 mg per mg of protein and combined type B adrenal cortex receptor Bmax is 294.5 ± 20.4 per mg protein

Page 35 1228988 V. Description of the invention (32) finol. The Kd value of [3H] prazosin binding to rat membrane proteins was 0.25 ± 0.01 nM, and the Bmax value of [3H] prazosin binding to rat membrane proteins was 71.3 to 1 / 7finol per milligram of protein. Calculate Ki with Ki = IC50 / (l + ([3H] ligand) / Kd), and the negative log result of pKi is Ki. The possibility is to exclude the possibility of interference analysis and evaluation of isoeugenodilol and other compounds. The compound was directly added to malondiacetaldehyde (diethyl acetal) (MDA, before the addition of thiobarbituric acid (TBA, Thiobarbituric Acid)).

Malonaldehyde bis (Diethyl Acetal)) standard. The results showed that in thiobarbituric acid-reactant (TBARS) analysis, isoeugenol diol (isoeugenodil 0115) had no effect on other compounds. As shown in Table 5, 'isoeugenol diol (15) and other compounds exhibit a lipid peroxidation response in a homogeneous state in the rat brain. Isoeugenol (15) dose-dose-ckpendently inhibits iron ion_ascorbic acid (Fe2 + _ascrbic⑽⑹)

Page 36 1228988 V. Explanation of the invention (33) The homogeneous state of rat brain induced by lipid peroxidation is 0 74 ± 0 · 03 μM (η = 6). In Table 3, 16 kinds of derivatives are more effective than ascorbic acid compared with compounds known to inhibit lipid peroxidation, and isoeugenol diol (15,15) and derivative 5 are ascorbic acid. Isoeugenol (15) inhibits lipid peroxidation, which is 3 times as much as α-vitamin 4α 4〇c〇pher〇1, 6 times ascorbic acid, and g of isosyring oil. (isoeUgen〇i) 11 times. Shame example 1: Synthesis of isoeugenodilol (15): Weigh 8 g of sodium hydroxide and dissolve in 100 ml of absolute ethanol, and then dissolve 164§ isoeugenol (1 mole) to dissolve In the above sodium hydroxide-containing ethanol solution, the solution was stirred and dissolved at room temperature. Then add 5 mol epichlorohydrin to react at room temperature, and determine whether the reaction is complete by thin layer chromatography (tlc). After the reaction was completed, the solution was concentrated by Lai, and the concentrated column was separated with a column filled with stone gum. Hexane: ethyl acetate 9 was used as the eluent. After the drops were collected, the solution was concentrated under reduced pressure to a supersaturated state, and then allowed to stand. White crude crystals were obtained. Repeated recrystallization with hexane to obtain purified white crystals, clove oil

Page 37 1228988 V. Description of the invention (34)

Phenol peroxide. Eugenol epoxide, its structure [4- (2,3_epoxypropoxy) _3_methoxy] small propenylbenzene) 〇 (2,3 · epoxypropoxy) _3 The molecular formula of -methoxy] -l-propylylbenzene) is C13H1603, and the molecular weight obtained by analysis by mass spectrometer is 220. ^ -NMRCCDCy 5 ··· 1.85-1.89 (d, 3H, Ar-CH = CH-CH3), 2J6-2.92 (m, 2H, Ar-0-CH2-HC-CH2_), 3.34-3.42 (m, 1H, Ar_0_CH2_HC_CHr), 3.88 (s, 3H, Ar-0-CH3), 4.05-4.09 (t, 4H, Ar-0-CH2-), 6.02-6.41 (m '1H' Ar-CH = CH-CH3) '6.84 6.89 (m, 3H, Ar), 7.26 (s, 1H, Ar-CH = CH-CH3). MSm / s: 220 (Scan EΓ). (2) Take 132g eugenol peroxide (0.6 mol) and 002g guaiacolethylamine (0.6 mol) in 100 ml of ethanol and heat Amination is carried out at 40-45 ° C. Stir overnight and let stand

The solid white crystals ‘after recrystallization from methanol’ can be used to obtain purified compounds. The structure of isoeugenodilol is {4_ [2-Cyclo-3_ (2_methoxy-1-oxoethylaminophenyl) propoxy] -3-fluorenyl M • propenylbenzene ( (4_``2_

Page 28 1228988 V. Description of the invention (35) The molecular formula of hydroxy-3- (2-methoxy-1 -oxyethylaminobenzene) propoxy] -3-methoxy} -l-propylenyl benzene) is c22H2905N, and the molecular weight obtained by analysis by mass spectrometer is 387 . Its argon nuclear magnetic resonance spectrum ih-NMR (CDC13) shows that 1.85-1.88 (d, 3H, At-CH = CH-CH3), 2.15-2 · 22 (br, 1H,-ΝΗ ·), 2.83-2.98 (m, 2H, Ar-0-CHrCH (Η Η) -CHr NH-), 3.07-3J0 (t, 2H, Ar-O-CHrCH2-NH-), 3.84-3.85 (d, 6H, Ar 0-CH3X2), 4.03-4.04 (t, 4H, Ar-0-CHrCHr), 4.12-4.15 (m, 1H, Ar-O-CHrCH (OH) -CH2-NH-), 6.05_6.17 (m 1H, Ar-CH = CH-CH3), 6.30-6.36 (d, 1H, Ar_CH = CH-CH3), 6.84_6.92 (m, 6H, Ar). MSm / s: 387 (Scan Γ). Infrared spectrum analysis IR (KBr): Vc has absorption of hydroxyl (〇 羟基) at 3587cm · 1, absorption of amine (NH) at 3587cm · 1, and absorption of methyl (CH2 ·) at 2947cm · 1, There was absorption of carbonyl (CO) at 1611 and 1591 cm · 1, and absorption of phenyl at 800 cm · 1. Ultraviolet absorption UV (CH3OH) Uog ε): 219 (3.79) nm, 257 (3.71) nm. Example 2 Synthesis of isoeugenolol

Vm Ijl

Page 39 1228988

(1) 8 g of hydrogen-emulsified nano-bath was dissolved in an appropriate anhydrous alcohol, 1 mol of isoeugenol was dissolved in the above-mentioned alcoholic sodium hydroxide solution, and the mixture was placed in a mantle heater and stirred. At the same time, 5 mol of epichlorohydrin was added at room temperature overnight without reaction. After the reaction was completed, the pressure was reduced, and the "Chenxi '" was separated into a column filled with crushed gel, and a white crude crystal was obtained by extracting the benzene and benzene = 3: 7. Crystallize again

It has been calcined repeatedly and recrystallized 'to obtain purified white crystals.

(2) Then take 22_0g of white crystals (0.1 mol) and dissolve them in absolute ethanol, and perform aminate reaction at room temperature; add 3 times the molar number of isopropylamine at room temperature at room temperature. A mantle heater was stirred overnight. After that, the reaction solution was concentrated under reduced pressure to remove unreacted isopropyl'amine. The concentrated liquid is left to stand to obtain coarse crystals which are hit by a solid. After several recrystallization steps with hexane, purified isoeugenolol can be obtained. Example 3 Synthesis of Ferulinolol

Chapter 1228988

Add 1500 g of eugenol to a reaction flask containing 900 ml of ethanol, and add isomolar sodium oxide, and allow it to react for 1 hour at 70t, then add 5 times more moles. Epichlorhydrin was refluxed under the same conditions for 2 hours. After the reaction was completed, the solvent was concentrated and evaporated to dryness under reduced pressure. Take 150ml dissolved in anhydrous methanol, add 3 times more moles of guaiacol ethylpropanolamine, and react at reflux at 55 ° C for 2 hours.

The obtained solution was directly concentrated under reduced pressure, suction-dried, and then purified by a column, and then crystallized from hexane to obtain purified ferulan (Femlin 0101).

Example 1 Synthesis of Femlidilol (16) ⑴ Synthesis of Ferulic acid peroxide

Add 15g of ferulic acid (feruiic aci (j) to a reaction flask containing ethanol and add sodium hydroxide of equal moles, and allow it to react at 70 ° C for 1 hour, then add 5 times more moles of Epichlorohydrin is refluxed under the same conditions for 2 hours. After the reaction is completed, the solvent is concentrated under reduced pressure to dry the solvent.

1228988 on page 41 'Explanation of the invention (38) is the eluent, the white crystal product ferulic acid peroxide (Ferulic acid peroxide) ° (2) Synthesis of guaiacoiethylpropanolamine (guaiacoiethylamine) a · 2 (2-methoxyphenoxy) ethyl bromide (2 (2_

Methoxyphenoxy) ethylbromide) Synthesis 22.4 ml guaiacol

(guaiacol '0.2 mole) and 34.6 ml ethylene dibromide (0.4 mole), mix and heat to i0 (TC, stir vigorously within 30 minutes, add i25 ml of 1.6N sodium hydroxide solution ， Continue heating and stirring until the pH reaches neutrality. The cooled mixture is extracted with chloroform. The organic layer is rinsed with 2N sodium hydroxide, then washed with a saturated sodium chloride solution and magnesium sulfate. After concentration, it is filled with silicone gel. The column was separated and an equal proportion of a mixed solvent of hexane and ethyl acetate was used as an eluent to obtain a white crystalline product.

b. [2 (2-methoxyphenoxy) ethyl] phthalimidine ([2 points

Methoxyphenoxy) ethyl] phthalimide) Synthesis

1228988 on page 42 —-—_________— V. Description of the invention (39) Take 36 g of 2 (2-Methoxyphenoxy) ethyl_bromide (0.156 mol) and 27.3 g Phthalimide (0.186 mol) was dissolved in 100 ml of dimethylacetamide, and heated to reflux at 90 ° C with stirring. 10.45 g of potassium hydroxide (0.85 mol) was dissolved in 30 ml of methanol to prepare a mixed solution. After 30 minutes, add the mixture and continue heating under reflux for 15 hours. The cooled mixture is poured into 300 ml of water, and the slowly precipitated solid is filtered. The filtered solid is added to a 200 ml 110% potassium carbonate (k2CO3) solution. , Heated and stirred vigorously for 10 minutes, the obtained mud was filtered, washed several times with water and recrystallized from anhydrous alcohol to obtain a white crystalline product. c · 2〇methoxyphenoxy) ethylamine (2- (2_

Methoxyphenoxy) ethylamine) Synthesis Take 21 g of [2 (2-methoxyphenoxy) ethyl] pyrimine ([2- (2-Methoxy

1228988, description of the invention (40) (hydrazine hydrate (0.071 mol)) was dissolved in 70 ml of absolute alcohol, heated to reflux for 45 minutes, 20 ml of 8% hydrochloric acid solution was added to the mixture, and the mixture was refluxed for 1 hour. The filtrate was filtered through cold The residue obtained by concentration under reduced pressure was basified with a 20% sodium hydroxide solution. The mixed solution was extracted with chloroform and dried with potassium carbonate. The filtrate was concentrated under reduced pressure. The concentrated solution was separated with a silica-filled column and ethyl acetate. For the eluent, a concentrated product was obtained. (3) Synthesis of Femlidilol Take 6.6 g of ferulic epoxide (ferulic epoxide, 0.03 and 50 g), and dissolve guaiacolethylamine (0.03 mol) in 30 In ml of absolute alcohol, stir at room temperature for 2 hours. The mixture is concentrated under reduced pressure and separated by a silica gel-filled column. The solvent is extracted with a mixture of hexane and ethyl acetate as the eluent. The final product, fermidilol (Femlidilol, 16), was obtained by recrystallization from hexane. Ferulidilol has a structure of i _ [(4_acrylic ethyl acetate_2 · methoxy) phenoxy] 3 [(2methoxymethoxyoxyethyl) amino] _propanol ethyl ester 2 methoxy) phenoxy] -3 [(2-methoxyphenoxyethyl) a

Page 44 1228988 V. Explanation of the invention (41) mineo] -propan〇i) The molecular formula is C24h3iN04, and the molecular weight measured by mass spectrometer is 446. Proton nuclear magnetic resonance spectroscopy CH-NMR, CDC13) 5 1.34 (t, 3H,-

COOCH2CH3), 2.93-3 · 18 (ιη, 4H, _CH2NHCH2), 3.77_3.84 (m, 6H, -ArOCH3), 4.06-4 · 14 (m, 4H,-ArOCH2), 4.17-4.20 (m , 3H, -COOCH2CH3), 4.24-4 · 31 (m, 1H, -CH (OH)), 6.26-6 · 34 (d, 1H, Ar-CH di-CH), 6.86-7 04 (m, 7H, Ar-H), 7.57-7.65 (d, 1H, Ar_CH = CH-). In the infrared spectrum ni (KBr) analysis, it was found that 3300 cm1 had an -NH- group absorption, and 1700 cm-1 had an carbonyl (-COOR) group absorption.

Page 45 1228988 V. Description of the invention (42)

0H

Table compound r2 Molecular formula 7 -CH = CHCOCH, -CH (CH〇7 c with NOa 8 -CH = CHCOCH, -C (CH 丄 C ^ H ^ NO, 9 -CH? CH, COCH, -C (CH,) , c17h? 7no, 10 -CHO -C (CH, h C ^ H ^ NO, 11 -ch, ch = ch2 -CH (CH,) 7 C ^ H ^ NO, 12 -CH = CHCH, -CH (CH丄 dNO; 13 -CH = CHCOOEt -C (CH 丄 14 CH2CH = CH2 -CH2CH2 〇 (C6H4) -2-OCH, c22h29no5 15 -ch = chch3 -CH2CH2 〇 (C6H4). 2-OCH, ^ 22 ^ 29 ^ 〇5 16 -CH = CHCOOEt -ch2ch2 〇 (c6h4). 2-OCH, c24h31no7

1ΪΗ Page 46 1228988 V. Description of the Invention (43) ΗΧΟ

2 Table compound Ri common name 1 -ch = chcoch3 Dehydrozingerone 2 -ch2ch2coch3 zingerone 3 -CHO Vanillin 4 -ch2ch = ch2 Eugenol 5 -ch = chch3 Isoeugenol 6 -CH = CHCOOH Ferulic acid

Page 47 1228988 V. Description of the invention (44) Table 3 β: «ι pAj compound right atrium 7.57 soil 0.09 7.68 soil 0.06 7.50 soil 0.007 left atrium 7.42 soil 0.08 8 · 53 ± 0 · 02 7 · 62 ± 0 · 09 ρΑ2 value trachea 6.93 ± 0.04 6.76 ± 0 · 05 6.77 ± 0.05 ρΑ2 value aorta < 5.00 < 5.00 < 5.00 10 11 12 13 14 15 16 (propran -olol) Labetal -ol Yateno (aten- 7.67 soil 0.03 8.23 soil 0.04 7.63 soil 0.008 7.62 soil 0.005 7.88 soil 〇12 7 · 83 ± 〇 · 12 8 · 04 Soil 〇 · 〇9 8 · 24 Soil 〇 · 〇6 7.91 Soil 〇09 · 7 · 89 Soil 0 · 11 8 · 36 ± 0 · 13 7 · 89 ± 0 · 12 7 · 54 ± 0 · 07 7 · 52 ± 0 · 05 7 · 80 ± 0 · 09 8 · 03 ± 0 · 15 8 · 07 ± 0 · 12 7 · 54 ± 0 · 16 7 · 66 ± 0 · 15 8 · 18 Soil 0 · 12 6 · 12 ± 0 · 05 6 · 28 ± 0 · 11 7 · 33 ± 0.15 7 · 76 ± 0.11 7 · 51 ± 0 · 06 8 · 07 ± 0 · 12 7 · 54 ± 0 · 16 < 5.00 < 5.00 < 5.00 < 5.00 7 · 05 ± 0.03 7 · 47 ± 0 · 45 7 · 05 ± 0 · 03 < 5.00 1.5 6 · 87 ± 0 · 08 2.3 7.34 soil 〇 · 〇3 5 · 70 ± 0 · 06 5 · 81 ± 0 · 06 < 5. 00 34.7 Table 4 Ventricle (β l) Lung (heart) m (a) — Ratio ratio β-blocker-log IC50 (M) pK, log IC50 (M) pKt log IC50 (M) pK isoeugenol double Isoeugenodilol 6.50 ± 0.07 7.36 6.84 ± 0.05 7.27 7.016 ± 0.06 7.41 Labetalol 7.52 ± 0.10 8.39 7.28 ± 0.04 7.83 7.03 ± 0.05 7.28 ----3.6 Atenolol 5.72 ± 0.04 6.58 4.53 ± 0.01 5.07 NT NT 32.4 — Proteanolol 8.77 ± 0.09 9.64 8.72 ± 0.11 9.26 NT NT 2.4 —

Page 48 1228988 V. Description of the invention (45) Table 5 Compound ic50 (μM) Compound IC50_) 1 35.7 2 28.2 3 6.94 4 12.8 5 1.52 6 6.29 7 83.4 _8 31.1 9 103.0 10 15.7 11 67.4 12 35.8 13 31.8 14 55.1 Isoeugenodilol (15) 0.74 ± 0.03 ferulidilol (16) 7.05 Ascorbic acid 4.10 ± 0.19 Trolox 3.04 ± 0.03 Vitamin E (a-Tocopherol) 1.83 ± 0.52 Table 6 Compounds Ri r2 mp ·, ° C Yield% Molecular formula 7 CH = CHCOCH, CH (CH,)? 109-111 55 c17h ?, no4 8 CH = CHCOCH, C (CH,), 96-98 35 c18h77no4 9 CH.CH .COCH, C (CH,), 75-77 30 c17h, 7no4 10 CHO C (CH,), 135-137 50 dN04 11 CH, CH = CH, CH (CH,) 7 42-44 68 c 12 CHNCHCH, CH (CH,) 7 120-122 31 dNO, M 13 CH = CHCOOEt C (CH,)? 149-151 45 C1QH? GNO, stomach 14 CH2CH = CH2 ch2ch2o (c6h4)-2-OCH, 46- 48 25 c22h29no5 15 ch = chch3 CH2CH20 (C6H4)-2-OCH, 125-127 52 c22h29no5 16 CH = CHCOOEt CH2CH20 (C6H4) -2 · 〇CH, 167-169 37 c24h31no7 _ i Page 49 1228988 The description of the list: Phenol derivative wooden structure table structure Table 2 Compound 3 pA2 values of 1-6, an acetate / ethylene type selectivity ratio

Table 4 Isoeugenodilol binding affinity-log 1C values Table 5 IC5 (3 inhibitory effect on lipid peroxidation Table 6 Physical data of 7-16 compounds Graphical illustration: Figure 1 7-16 compounds Synthetic method Figure 2 Isoeugenol blood pressure and heartbeat dose (#) 0.1 mk / kg (V) 0.5 mk / kg (T) 1.0 mk / kg (□) 3.0 mk / kg (〇) Control group

Page 28 1228988 Schematic illustration Figure 3 Isoeugenol blood pressure and heartbeat dose (#) 10 mk / kg (V) 30 mk / kg (▼) 100 mk / kg (〇) Control group Figure 4 K + induction Aortic contraction (A) High K + concentration (□) Before adding Bay K 8644 () After adding Bay K 8644 (B) Bay K 8644 (□) Adding tetraethylammonium vaporized (TEA) () Add 1CT6M glibenclamide Figure 5 Ca + 2 induced aortic contraction (□) EGTA physiological solution (CaCl2) Figure 6 Inhibition of norepinephrine or hydroxychrome-induced contraction

1RH P.51 1228988 Brief description of the diagram (A) norepinephrine (B) serotonin (〇) control group isoeugenodilol dose (φ) IO-8 M,

(▽) 10-7M, (▼) 10.6M, () 10-5M Figure 7 Atrial and tracheal effects (A) Right atrial positive change time (B) Left atrial positive change time (C) Tracheal relaxation Isoeugenodilol dose (#) ΙΟ 8 $ (▽) 10.7M, (T) 10.6M? (〇) Control group Figure 8 Atrial and tracheal effects (Α) Positive right atrium change Temporal (B) Positive left atrial change (C) Tracheal relaxation

Page 52 1228988 Schematic description (#) L-isoproterenol (〇) Control group (▽) propranolol (▼) isoeugenodilol

Page 53

Claims (1)

1228988 Case No. 89118397 Amendment VI. Linearity C3v of unsaturated groups in the scope of patent application and < 丨 / 〇〇⇨5'1 ^ may be (: 3_6,-(^ 60 (〇 凡) -2- 0 (: 丨 _3 -CuOCQE ^ -OCw, -CmCXQ ^ M-OCw. OH / * Formula π 4. A pharmaceutical composition with anti-oxidant activity, which is mainly composed of derivatives of formula II with guaiacoxypropanolamine and vanilloid structure, and various excipients are added if necessary. Or diluent; its name may be -C ^ COQ-5, linear c 3-6 'and -Cj_5COOC containing unsaturated group 5, R2 may be C3-6' -C 丨 _60 (C ( sH4) -2-〇C 丨 · 3, -C 丨 · 60 (^ 6Η4) -3-〇 (^ Bu 3, -C | _6〇 (C6H4) -4-OCj_3. 5. A kind of derivative with a Vanilloid structure, which is a compound having the main structure of Formula II. Ri may be -C3-6C0Ci_5, linear C3v containing unsaturated groups, and 4-5. ) 0 (: 1-5, ruler 2 can be C3-6, Page 55 1228988 £ ^ 89118397 VI. Scope of patent application Formula II 6.—A method for manufacturing a compound of formula η having a Vanilloid structure. A derivative of the structure I of Formula I, whose R 丨 can be -C3-6COCM, a linear chain c3_6 containing an unsaturated group, and- CmCOOC 丨 _5 For raw materials', Propanolamine is obtained by reacting with epichlorohydrin under test environment, and then separately with isopropyl amine, tert-butylamine or guaiacoxyethylamine (guaiacoxyethylamine) reaction to obtain, for example, Ri may be -CwCOCu, linear C3_6 containing unsaturated groups, and CwCOOCm, R2 may be C3.6, -CmOCQHOIOCm, -C 丨 _60 (C6H4)-3-OCio, as shown , A compound with a Vanilloid structure. Formula II Page 56