JP2006517910A - New α-glucosidase inhibitors derived from natural sources - Google Patents

Abstract

The present invention can be applied to the treatment of diabetes, cancer, and other viral diseases such as hepatitis B / C, HIV, AIDS, and the like, which can be applied to pipetalin (Formula 1a), sesamin (Formula 1b), peritrine (Formula 1c), guanine ( The present invention relates to a method for providing an α-glucosidase inhibitory effect to a patient by administering a pharmaceutical composition comprising a formula 1d), an α-glucosidase inhibitor selected from brachystamide-B (formula 1e) Furthermore, this invention provides the method of isolate | separating (alpha) glucosidase inhibitor with the high yield from the Indian root pepper which is a plant source.

Description

  The present invention relates to pipeataline (formula 1a), sesamin (formula 1b), peritorine (formula 1c), guineensine (formula 1d), brachystamide-B (formula 1b) It relates to a method of providing an α-glucosidase inhibitory effect to a patient by administering a pharmaceutical composition having an α-glucosidase inhibitor selected from 1e). Specifically, the present invention relates to five compounds from the plant source, Piper longum, namely, pipeataline [5- (1-dodecenyl) -1,3-benzodioxole], sesamin [5. , 5- (Tetrahydro-1H, 3H-furo (3,4-e) furan-1,4-diyl) bis-1,3-benzodioxole], peritrine [N- (2-methylpropyl) -2 , 4-Decaazienamide], Guineansin [13- (1,3-benzodioxol-5-yl) -N (2-methylpropyl) -2,4,12-tridecatrienamide], Braxamide-B [[ 15- (1,3-benzodioxol-5-yl) -N (2-methylpropyl) -2,4,14-pentadecatrienamide] with high yield. In addition, the present invention applies these compounds to treatment as α-glucosidase inhibitors in the form of pharmaceutical preparations suitable for diabetes, cancer, and other viral diseases such as hepatitis B / C, HIV, AIDS, etc. Clarify how to do.

  The use of plants as drugs dates back to ancient times. In fact, there remains evidence that ancient Indian, ancient Chinese and ancient North African civilizations have used plants effectively to treat a wide range of diseases (Phillipson JD, phytochemistry, 2001, 56, 237-243). Later, as the knowledge spread through new research and clinical experience, changes in pharmacotherapy have also been seen. With the development of new diseases and the identification of targets that can be applied to various treatments, the search for new compounds with unique structures and properties is in progress.

  The α-glucosidase enzyme is recognized as such a target. Metabolic abnormalities such as diabetes, obesity, type IV hyperlipoproteinemia (Trusch eit, E. et al., Angew. Chem .. Int. Ed., Engl., 1981, 20, 744-761; Puls, W. Keupu, Diabelologia, 1973, 9, 97; Puls, W Habilitationssoh, Chrift universitat Dusseldorf, 1980), HIV, hepatitis B virus, human cytomegalovirus, influenza (Heightman TD and Vasella AT, Angew. Chem. Int. Ed. , Engl., 1999, 38, 750-770; Mehta et al., FEBS Lett., 1998, 430, 17-22; Watson AA et al., Phytochemistry, 2001, 56, 265-295), cancer, immunodeficiency In the state, methods for applying inhibitors of a-glucosidase to therapy are being found one after another. Blood levels of glucosidase have been found to increase in many patients with various tumors (Woollen, JW and Tesiar, P., 1965, din. Chem. Ada., 12, 671-683) It is understood to be involved in the degradation of extracellular matrix during cell invasion (Bernaki, RJ et al., 1985, “Cancer metastasis Rev 4”, 81-102). Therefore, degrading glucosidase inhibitors have been actively studied as cancer treatments (Watson, A. A. et al., Phytochemistry, 2001, 56, 265-295).

  Various compounds that have the ability to inhibit a-glucosidase have already been investigated for usefulness in chemotherapy (El Ashry et al., Pharmazie, 2000, 55, 251-262, 331-348, 403-415) . a. Some drugs aimed at inhibiting glucosidase are used in clinical therapy or are in various clinical development stages (Drugs of the future, 1986, 11, 795-797; Drugs of the future 1986, 11, 1039-1042; Watson AA et al., Phytochemistry, 2001, 56, 265-295), the magnitude of the mental burden of the illnesses described above indicates a clear need for new drugs. There is also a need to secure a large pool of inhibitors because patients may acquire resistance to current therapies.

  In the past, traditional medical practices and knowledge gained from screening of extracts from plants and animals has created new natural products as bioactive agents with the potential to treat human diseases. (Gullo. VP, The discovery of natural products with therapeutic potential, Butterworth-Heinemann, Boston, 1994; Cragg G. Metal J., Nat. Prod., 1997, 60: 52-60).

  By screening natural sources, a number of clinically useful drugs have been discovered that play important roles not only in the treatment of the diseases mentioned above but also in the prevention of such diseases. Similar to increased drug resistance, the clinical significance of the prevalence of diabetes, cancer, HIV and other viral diseases is increasing, identifying new sources of active compounds, There is a growing urgency to establish a large pool of such compounds.

  As will be described later, as a result of searching for a-glucosidase inhibitors from conventional medicinal plants, it was found that Indian pepper has a high yield of potent a-glucosidase inhibitors.

  Indian persimmon (Piper lingum Linn.) Has been described in traditional Indian medicine as effective for malaria fever, heart disease, splenomegaly, cough, edema (PV Sharma) , Classical uses of medicinal plants, Haridas Ayurveda series (4), Chaukambha Viswabharathi, Varanasi, 1996).

  The present invention provides potent aglucosidase inhibitors in the form of suitable pharmaceutical formulations that can be applied therapeutically in the treatment of diabetes, cancer, tumors, metastasis, immunomodulation and as a broad range of antiviral agents. It relates to identification and isolation from Indian pepper.

  Table 1 shows each species of the genus Pepper from which the compounds claimed as a-glucosidase inhibitors in the present invention are obtained. Table 2 shows their biological activity.

(Application and administration)
The α-glucosidase inhibitor of the present invention includes diabetes, cancer, HIV, AIDS, hepatitis B or C, other viral infections, immunodeficiency states, multiple sclerosis, which are improved and cured by inhibition of α-glucosidase, It can be applied or administered by conventional treatments and therapies for arthritis and the like.

  When applied to humans, animals and / or livestock, the compounds as a-glucosidase inhibitors of the present invention can be administered through various routes depending on the suitability and clinical condition. When applied to humans, compounds as alpha glucosidase inhibitors can be administered through various routes, such as oral, intraperitoneal, intravenous, and / or intramuscular injection, depending on the condition.

(Formulation)
a. Compounds of the invention as glucosidase inhibitors should be formulated using any pharmaceutically applicable additive, carrier, excipient that will in no way alter the efficacy and properties of the compound. Can do.

  When applied to humans, the compounds of the present invention as alpha glucosidase inhibitors may be formulated using a number of carriers and additives well known in the art that are useful for pharmaceutical administration and pharmaceutically acceptable. Can do.

  The selected carrier or excipient will of course be compatible with the mode of application or administration of the glucosidase inhibitor.

(Effective level)
The expression “effective amount” and / or “suppressing amount” refers to the amount of the a-glucosidase inhibitor compound of the present invention that occurs in an untreated control group under an appropriate therapeutic condition depending on the disease state and severity. Compared to, for example, to reduce the degree of illness, such as a decrease in postprandial blood glucose and insulin levels in the case of diabetes, or, in some cases, to significantly reduce cancer, tumors, or viral infections Means the amount considered. This implies that an effective amount of an a-glucosidase inhibitor compound of the present invention is less than an amount that produces undesirably significant side effects in a living organism being treated for a particular disease. This was supplemented by using the expression “pharmaceutically effective amount”. The actual application rate and amount will vary depending on the disease state. This is independent of concentration as illustrated in the examples of the present invention.

  The actual rate and amount of application will vary depending on the severity of the disease and / or infection and will also depend on several factors, such as the age and sex of the individual being treated and the mode of administration. Those having ordinary skill in the art can readily determine the actual dose and dosing regimen taking these factors into account.

コショウ属由来の化合物は、幅広い生物活性を示す。表２は、ピパタリン、セサミン、ペリトリン、ギネアンシン、およびブラキスタミド−Ｂの生物活性を示している。

Compounds from the genus Pepper show a wide range of biological activities. Table 2 shows the biological activity of pipetalin, sesamin, peritrine, gineansin, and brachystamide-B.

  The main object of the present invention is to provide a new activity of pipetalin, sesamin, peritrine, gineansin, or brachystamide-B obtained as an α-glucosidase inhibitor from Indian pepper.

  Another object of the present invention is to provide a method of treating a patient to inhibit alpha glucosidase.

  Another object of the present invention is in the treatment and treatment of human illnesses such as hyperglycemia, hyperinsulinemia, hyperlipoproteinemia, cancer, viral infection, hepatitis B / C, HIV, AIDS, etc. It relates to applying the above compounds as therapeutics as α-glucosidase inhibitors.

  Another object of the present invention provides a method of treating a patient to inhibit alpha-glucosidase using a pharmaceutical agent having pipetatin, sesamin, peritrine, gineansin, or brachystamide-B obtained from Indian pepper. That is.

  Furthermore, the object of the invention relates to the separation of pipetalin from completely new raw materials.

  Yet another object of the present invention relates to the separation of five compounds from Indian pepper, namely, pipataline, sesamin, peritrine, gineansin, braxamide-B.

  Still another object of the present invention is to provide a method for separating pipeatalin, sesamin, peritrine, guaneancin, or brachystamide-B obtained from Indian pepper with good yield.

  Accordingly, the present invention relates to a pharmaceutical composition comprising an alpha glucosidase inhibitor selected from pipetalin (formula 1a), sesamin (formula 1b), peritrine (formula 1c), guaneancin (formula 1d), brachystamide-B (formula 1e). To provide a patient with an α-glucosidase inhibitory effect.

  The present invention is an α-glucosidase inhibitor in the treatment and treatment of human diseases such as hyperglycemia, hyperinsulinemia, hyperlipoproteinemia, cancer, viral infection, hepatitis B / C, HIV, AIDS, etc. , New activity of pipetalin, sesamin, peritrine, gineansin, or brachystamide-B obtained from Indian pepper.

  The invention also relates to the separation of a new raw material, i.e. pipetalin from Indian pepper. Another aspect of the present invention is to provide a method for separating pipeatalin, sesamin, peritrine, gineansin, or brachystamide-B obtained from Indian pepper with good yield.

  According to the object of the present invention, the present invention provides a method for providing an α-glucosidase inhibitory effect to a patient, comprising hyperglycemia, hyperinsulinemia, hyperlipoproteinemia, cancer, viral infection, type B in the patient. And in the treatment and treatment of diseases such as hepatitis C, HIV, AIDS, etc., selected from pipetalin (Formula 1a), sesamin (Formula 1b), peritrine (Formula 1c), guaneancin (Formula 1d), braxamide-B (Formula 1e) A method comprising: administering to a patient an effective amount of a pharmaceutical formulation having the α-glucosidase inhibitor to be administered and a pharmaceutically acceptable ingredient.

In one embodiment of the invention, pipataline provides α-glucosidase inhibitory activity of up to 77.45% with an IC ₅₀ value of 26.52 (μg / ml).

In another embodiment of the invention, sesamin provides α-glucosidase inhibitory activity up to 76.18% with an IC ₅₀ value of 36.35 (μg / ml).

In another embodiment of the invention, peritrine provides up to 86.03% alpha glucosidase inhibitory activity with an IC ₅₀ value of 34.43 (μg / ml).

In another embodiment of the invention, the guaneancin provides up to 61.71% α-glucosidase inhibitory activity with an IC ₅₀ value of 20.15 (μg / ml).

In another embodiment of the invention, brachystamide-B provides up to 73.90% α-glucosidase inhibitory activity with an IC ₅₀ value of 33.61 (μg / ml).

  In another embodiment of the invention, a pharmaceutical formulation comprising pipetalin, sesamin, peritrine, guaneancin, or brachystamide-B optionally comprises a pharmaceutically acceptable ingredient.

  Yet another embodiment of the present invention provides for the first time a method of separating pipetalin from Indian pepper.

In a further embodiment of the present invention, from the plant source Indian Pepper, selected from pipetalin (Formula 1a), sesamin (Formula 1b), peritrine (Formula 1c), guaneancin (Formula 1d), braxamide-B (Formula 1e) A method for isolating the alpha glucosidase inhibitor, comprising:
a. Extracting from a dried fruit of Indian pepper using a solvent;
b. Concentrating the extract under vacuum to obtain a residue;
c. Eluting the residue of step (b) using hexane to obtain pipetalin and residue;
d. Eluting the residue of step (c) using hexane containing about 3% ethyl acetate to obtain sesamin and residue;
e. Eluting the residue of step (d) using hexane containing about 5% ethyl acetate to obtain peritrine and residue;
f. Eluting the residue of step (e) using hexane containing about 10% ethyl acetate to obtain guaneancin and residue;
g. Eluting the residue of step (f) using hexane containing about 11% ethyl acetate to obtain brachystamide-B;
Is included.

  In one embodiment, the solvent used in step (a) is selected from hexane, cyclohexane, n-pentane.

  Another embodiment of the invention relates to the separation of pipetalin from completely new raw materials.

  Yet another embodiment of the invention relates to the separation of these compounds from Indian pepper as o-glucosidase inhibitors.

  The present invention embodies the separation of pipataline, sesamin, peritrine, gineansin, brachystamide-B as o-glucosidase-inhibiting components from Indian Pepper, of which pipataline is from a completely new raw material.

  The present invention relates to five types of compounds from Indian plant pepper, ie, pipataline [5- (1-dodecenyl) -1,3-benzodioxole], sesamin [5,5- (tetrahydro-1H, 3H-furo (3,4-e) furan-1,4-diyl) bis-1,3-benzodioxole], peritrine [N- (2-methylpropyl) -2,4-decadienamide], ginanthin [ 13- (1,3-benzodioxol-5-yl) -N (2-methylpropyl) -2,4,12-tridecatrienamide], braxamide-B [[15- (1,3-benzo [Dioxol-5-yl) -N (2-methylpropyl) -2,4,14-pentadecatrienamide] in high yields. The invention also relates to new uses of these compounds as a-glucosidase inhibitors.

  The present invention embodies the separation of five types of a-glucosidase inhibitory components, pipataline, sesamin, peritrine, gineansin, and brachystamide-B from Indian pepper, of which pipataline is from a completely new raw material.

  Some of the embodiments of the present invention are represented by the following examples. These examples are not to be considered as limiting the scope of the invention.

In another embodiment of the present invention, pipetalin obtained from Indian pepper has the following spectrochemical and physical properties.
Molecular Formula: C ₁₉ H ₂₈ O ₂
MP: 38 ° C.
IR (KBr) γ _max cm- ¹
2829,1468,1248,1040,980,960.
'H NMR (200 MHz, CDCI ₃ ) (δ)
0.95 (3H, t, Hl), 1.20-1.60 (16H, b, H-2-9), 2.20 (2H, q, H-10), 5.90 (2H, s, -OC H ₂ O), 6.0-6.15 (1H, q, Hl 1), 6.30 (1H, d, J = 15.5 Hz, H-12), 6.70 (2H, s, H-5 ', 6'), 6.88 (1H, s, H -2 ¹ ).
^I3 C NMR (50 MHz, CDC1 ₃ )
14.12 (Cl), 22.71 (C-2), 29.37-29.66 (C-3-8), 31.95 (C-9), 32.95 (C-10), 100.89 (-), 105.46 (C-20, 108.20 (C-5 ¹ ), 120.17 (C-6 ^f ), 129.27 (C-ll), 129.57 (Cl 2), 132.61 (C-1 '), 146.68 (C-4'), 148.01 (C-3 ' )
EI-MS
M ⁺ 288

In another embodiment of the invention, sesamin has the following spectrochemical and physical properties:
Molecular Formula: C ₂₀ H ₁₈ O ₆
MP: 122 ° C
UV λ _max (EtOH)
286, 235nm
IR (KBr) γ _max cm ^-1
2800, 1600, 1071, 925, 913, 718 cm ^-1
'H NMR (200 MHz, CDC1 ₃ ) (δ):
3.08 (IH, m, H-8), 3.90 (IH, dd, J = l0Hz, 4Hz, H-2b), 4.20-4.30 (IH, m, H-2a), 4.75 (IH, d, J = 5Hz , H-4), 6.0 (2H, s, -OC H ₂ O-), 6.80 (2H, s, H-2 ', 5% 6.84 (IH, s, H-6')
EI-MS
^{- M + 354, 203, 161,149} .
[a] _D + 78.3 °

In another embodiment of the invention, peritrine has the following spectrochemical and physical properties:
Molecular formula: C ₁₄ H ₂ sNO
MP: 83 ° C.
UV λmax (EtOH)
260nm.
IR (KBr) γ _max cm ^-1
3260, 1655, 1600, 1255 cm ^-1 .
H NMR (200 MHz, CDCl ₃ ) (δ)
0.91 (6H, d, J = 6Hz), 0.8-1.0 (3H), 1.25 (6H, bs), 1.7-2.4 (3H, m), 3.15 (2H, t), 5.55 (IH, t), 5.75 ( IH, d, J = 15 Hz), 6.0-6.2 (2H, m), 6.80-7.20 (IH, m).
EI-MS m / z (%)
223 (M ⁺ , 33), 208 (7), 180 (6), 166 (6), 152 (33), 151 (100), 96 (50), 81 (64), 72 (4), 57 ( 16), 43 (10).

In another embodiment of the present invention, guaneancin [13- (1,3-benzodioxol-5-yl) -N (2-methylpropyl) -2,4,12-tridecatrienamide] is Has the following spectrochemical and physical properties.
Molecular Formula: C ₂₄ H ₃₃ N0 ₃
MP: 119 ° C.
UV λmax (MeOH)
261 nm.
IR (KBr) Ymax cm- '
3300, 1655, 1630, 1545, 1250, 1035cm ' ¹ .
'H NMR (200 MHz, CDCl ₃ ) (δ)
0.93 (6H, d, J = 6.5 Hz), 1.25-1.50 (8H), 1.80 (IH, m), 2.12-2.21 (4H, m), 3.16 (2H, t, J = 6.4 Hz), 5.48 (IH , br), 5.74 (IH, d, J = 15.0Hz), 5.93 (2H, s), 6.05-6.15 (3H, m), 6.28 (IH, d, 15.5 Hz), 6.72-6.90 (3H), 7.19 (IH, dd, J = 15Hz, lOHz).
EI-MS
383 (M ⁺ , 35), 249 (32), 180 (22), 152 (45), 135 (100).

In another embodiment of the invention, brachystamide-B has the following spectrochemical and physical properties:
Molecular formula: C ₂₆ H ₃₇ NO ₃
UV λmax (EtOH)
260, 208nm.
IR (KBr) y _max
1654, 1625, 1000.
'H NMR (200 MHz, CDCI ₃ ) (δ)
0.86 (6H, d, H-3 ", 4"), 1.20-1.70 (12H, b, H-7-12), 1.70-1.90 (IH, m, H-2 '), 2.05-2.20 (2H, m, H-6, 15), 3.15 (2H, t, Hl "), 5.68 (IH, d, J = 15.5Hz, H-2), 5.84 (2H, s, -OC H ₂ O-), 5.95 -6.15 (3H, m, H-4, 5, 14), 6.23 (IH, d, J = 16Hz, H-15), 6.72 (2H, s, H-5 ', 6'), 6.83 (IH, s, H-2 \ 7.12 (IH, m, H-3).
¹³ C NMR (50 MHz, CDCI ₃ ) (6)
166.39 (C-1), 121.85 (C-2), 142.92 (C-3), 128.33 (C-4), 141.23 (C-5), 32.86 (C-6),
28.64-29.51 (C-7-12), 32.81 (C-13), 129.36 (C-14), 129.42 (C-15), 132.58 (C-1 ¹ ),
105.48 (C-2 '), 147.96 (C-3% 146.59 (C-40, 108.21 (C-5% 120.18 (C-60, 46.97 (C-1 ")),
28.66 (C-2 "), 20.69 (C-3", 4 "), 100.88 (-OC & O-).
EI-MS
M ⁺ 411, 396 (42), 299 (20), 149 (28), 97 (30), 69 (88), 57 (100).

Experimental Method: Method for Separating Pipatalin, Sesamin, Peritrine, Guineansin, Brachystamide-B Dried and powdered Indian Pepper fruit (500 g) was placed in a Soxhlet extractor. The powder was extracted with hexane, and the resulting hexane extract was concentrated under vacuum. The dark green residue was introduced into a silica gel column packed to 60-120 mesh, diameter 3.5 cm, height 60 cm.
First, elution from the column with hexane yielded pipetalin. The yield of pipetalin was about 6.0 g. Further, elution from the column using hexane containing 3% ethyl acetate gave sesamin. Sesamin yield was about 200 mg.
Further, elution was performed from the column using hexane containing 5% ethyl acetate to obtain peritrine. The yield of peritrine was about 200 mg.
Further, elution from the column using hexane containing 10% ethyl acetate gave guaneancin. The yield of guaneancin was about 300 mg.
Further, elution from the column using hexane containing 11% ethyl acetate gave Braxamide-B. The yield of brachystamide-B was about 120 mg.
All of the above compounds were obtained with a purity of 90%.
The spectrochemical and physical properties of the above compounds are as follows.

Pipatalin has the following spectrochemical and physical properties.
Molecular Formula: C ₁₉ H ₂₈ O ₂
MP: 38 ° C.
IR (KBr) γ _max cm- ¹
2829,1468,1248,1040,980,960.
'H NMR (200 MHz, CDCI ₃ ) (δ)
0.95 (3H, t, Hl), 1.20-1.60 (16H, b, H-2-9), 2.20 (2H, q, H-10), 5.90 (2H, s, -OC H ₂ O), 6.0-6.15 (1H, q, Hl 1), 6.30 (1H, d, J = 15.5 Hz, H-12), 6.70 (2H, s, H-5 ', 6'), 6.88 (1H, s, H -2 ¹ ).
^I3 C NMR (50 MHz, CDC1 ₃ )
14.12 (Cl), 22.71 (C-2), 29.37-29.66 (C-3-8), 31.95 (C-9), 32.95 (C-10), 100.89 (-), 105.46 (C-20, 108.20 (C-5 ¹ ), 120.17 (C-6 ^f ), 129.27 (C-ll), 129.57 (Cl 2), 132.61 (C-1 '), 146.68 (C-4'), 148.01 (C-3 ' )
EI-MS
M ⁺ 288

Sesamin has the following spectrochemical and physical properties.
Molecular Formula: C ₂₀ H ₁₈ O ₆
MP: 122 ° C
UV λ _max (EtOH)
286, 235nm
IR (KBr) γ _max cm ^-1
2800, 1600, 1071, 925, 913, 718 cm ^-1
'H NMR (200 MHz, CDC1 ₃ ) (δ):
3.08 (IH, m, H-8), 3.90 (IH, dd, J = l0Hz, 4Hz, H-2b), 4.20-4.30 (IH, m, H-2a), 4.75 (IH, d, J = 5Hz , H-4), 6.0 (2H, s, -OC H ₂ O-), 6.80 (2H, s, H-2 ', 5% 6.84 (IH, s, H-6')
EI-MS
^{- M + 354, 203, 161,149} .
[a] _D + 78.3 °

Peritrine has the following spectrochemical and physical properties.
Molecular formula: C ₁₄ H ₂ sNO
MP: 83 ° C.
UV λmax (EtOH)
260nm.
IR (KBr) γ _max cm ^-1
3260, 1655, 1600, 1255 cm ^-1 .
H NMR (200 MHz, CDCl ₃ ) (δ)
0.91 (6H, d, J = 6Hz), 0.8-1.0 (3H), 1.25 (6H, bs), 1.7-2.4 (3H, m), 3.15 (2H, t), 5.55 (IH, t), 5.75 ( IH, d, J = 15 Hz), 6.0-6.2 (2H, m), 6.80-7.20 (IH, m).
EI-MS m / z (%)
223 (M ⁺ , 33), 208 (7), 180 (6), 166 (6), 152 (33), 151 (100), 96 (50), 81 (64), 72 (4), 57
(16), 43 (10).

Guineansin [13- (1,3-benzodioxol-5-yl) -N (2-methylpropyl) -2,4,12-tridecatrienamide] has the following spectrochemical and physical properties: .
Molecular Formula: C ₂₄ H ₃₃ N0 ₃
MP: 119 ° C.
UV λmax (MeOH)
261 nm.
IR (KBr) Ymax cm- '
3300, 1655, 1630, 1545, 1250, 1035cm ' ¹ .
'H NMR (200 MHz, CDCl ₃ ) (δ)
0.93 (6H, d, J = 6.5 Hz), 1.25-1.50 (8H), 1.80 (IH, m), 2.12-2.21 (4H, m), 3.16 (2H, t, J = 6.4 Hz), 5.48 (IH , br), 5.74 (IH, d, J = 15.0Hz), 5.93 (2H, s), 6.05-6.15 (3H, m), 6.28 (IH, d, 15.5 Hz), 6.72-6.90 (3H), 7.19 (IH, dd, J = 15Hz, lOHz).
EI-MS
383 (M ⁺ , 35), 249 (32), 180 (22), 152 (45), 135 (100).

Brachystamide-B has the following spectrochemical and physical properties.
Molecular formula: C ₂₆ H ₃₇ NO ₃
UV λmax (EtOH)
260, 208nm.
IR (KBr) y _max
1654, 1625, 1000.
'H NMR (200 MHz, CDCI ₃ ) (δ)
0.86 (6H, d, H-3 ", 4"), 1.20-1.70 (12H, b, H-7-12), 1.70-1.90 (IH, m, H-2 '), 2.05-2.20 (2H, m, H-6, 15), 3.15 (2H, t, Hl "), 5.68 (IH, d, J = 15.5Hz, H-2), 5.84 (2H, s, -OC H ₂ O-), 5.95 -6.15 (3H, m, H-4, 5, 14), 6.23 (IH, d, J = 16Hz, H-15), 6.72 (2H, s, H-5 ', 6'), 6.83 (IH, s, H-2 \ 7.12 (IH, m, H-3).
¹³ C NMR (50 MHz, CDCI ₃ ) (6)
166.39 (C-1), 121.85 (C-2), 142.92 (C-3), 128.33 (C-4), 141.23 (C-5), 32.86 (C-6), 28.64-29.51 (C-7- 12), 32.81 (C-13), 129.36 (C-14), 129.42 (C-15), 132.58 (C-1 ¹ ), 105.48 (C-2 '), 147.96 (C-3% 146.59 (C- 40, 108.21 (C-5% 120.18 (C-60, 46.97 (C-1 "), 28.66 (C-2"), 20.69 (C-3 ", 4"), 100.88 (-OC & O-).
EI-MS
M ⁺ 411, 396 (42), 299 (20), 149 (28), 97 (30), 69 (88), 57 (100).

Quantification of α-Glucosidase Inhibitory Activity of Compounds Isolated from Indian Pepper An evaluation of α-glucosidase inhibition was performed by a color development method. In summary, 10 μl of test compound (5 mg / ml, subsequently diluted) dissolved in DMSO was incubated for 5 minutes with 5 μl of yeast α-glucosidase prepared in 100 mM phosphate buffer (pH 7.00). After 5 minutes incubation, 50 fil of 5 mM substrate (p-nitrophenyl-α-D-glucopyranoside prepared in the same buffer) was added. Absorbance was recorded at 405 nm by spectrophotometry before and 5 minutes after addition of substrate. It was observed that the absorbance increased after the reaction after the addition than before the substrate was added. Inhibition rates were calculated as (1-OD test compound / OD control compound) x 100 and 50% inhibitory concentration (IC50) was calculated by applying the appropriate regression analysis.

  In accordance with the practice of the present invention, it has been found that pipataline, sesamin, peritrine, gineansin, and brachystamide-B are isolated from Indian pepper, of which pipataline is from a completely new raw material. The yield of these compounds is also considerable. Furthermore, it has been found that all of these compounds exhibit α-glucosidase inhibitory properties.

(advantage)
Alpha-glucosidase inhibitors have recently attracted attention due to their broad activity in diseases of various causes, such as diabetes, viral diseases, cancer, HIV, hepatitis B / C and the like. At present, there is more interest in obtaining α-glucosidase inhibitors from natural sources.

  The compounds pipetalin, sesamin, peritrine, gineansin, brachystamide-B are used in pure form. Therefore, it is very important to separate from the present invention, i.e., Peptarin, sesamin, peritrine, gineansin, and brachystamide-B as α-glucosidase inhibitors at a high yield.

(A) chemical formula of pipetalin [5- (1-dodecenyl) -1,3-benzodioxole], (b) sesamin [5,5- (tetrahydro-1H, 3H-furo (3,4-e) furan -1,4-diyl) bis-1,3-benzodioxole], (c) peritrine [N- (2-methylpropyl) -2,4-decaazienamide], (d) ginanthin [13 -(1,3-benzodioxol-5-yl) -N (2-methylpropyl) -2,4,12-tridecatrienamide], (e) Brachystamide-B [[15- (1 , 3-Benzodioxol-5-yl) -N (2-methylpropyl) -2,4,14-pentadecatrienamide] Graph showing o-glucosidase inhibitory activity of pipetalin, sesamin, peritrine, gineansin, and brachystamide-B

Claims (14)

  In the treatment and treatment of illnesses such as hyperinsulinemia, hyperlipoproteinemia, viral infections, hepatitis B and C, HIV, etc. in patients, pipetalin (formula 1a), sesamin (formula 1b) and peritrine ( A pharmaceutical formulation having an effective amount of an alpha glucosidase inhibitor selected from the group consisting of Formula 1c), Gineanthin (Formula 1d), and Brachystamide-B (Formula 1e), and a pharmaceutically acceptable ingredient Of said pharmaceutical formulation by administering to said patient. 2. Use according to claim 1, wherein pipetalin provides α-glucosidase inhibitory activity up to 77.45% and an IC ₅₀ value of 26.52 (μg / ml). The use according to claim 1, wherein sesamin provides α-glucosidase inhibitory activity up to 76.18% and IC ₅₀ value is 36.35 (μg / ml). The use according to claim 1, wherein peritrine provides an α-glucosidase inhibitory activity of up to 86.03% and an IC ₅₀ value of 34.43 (μg / ml). The use according to claim 1, wherein guaneancin provides up to 61.71% α-glucosidase inhibitory activity and has an IC ₅₀ value of 20.15 (μg / ml). The use according to claim 1, wherein brachystamide-B provides α-glucosidase inhibitory activity up to 73.90% and has an IC ₅₀ value of 33.61 (μg / ml). From the group of Indian Naga pepper, which is a plant source, from the group consisting of pipetalin (Formula 1a), sesamin (Formula 1b), peritrine (Formula 1c), Guineansin (Formula 1d), and Brachystamide-B (Formula 1e) A method for isolating a selected α-glucosidase inhibitor comprising:
a) performing extraction using a solvent from the dried fruit of Indian Pepper;
b) concentrating the extract of step (a) under vacuum to obtain a residue;
c) eluting the residue of step (b) using hexane to obtain pipetalin and residue;
d) eluting the residue of step (c) using hexane containing about 3% ethyl acetate to obtain sesamin and residue;
e) eluting the residue of step (d) using hexane containing about 5% ethyl acetate to obtain peritrine and residue;
f) eluting the residue of step (e) using hexane containing about 10% ethyl acetate to obtain guaneancin and residue;
g) eluting the residue of step (f) using hexane containing about 11% ethyl acetate to obtain brachystamide-B;
Including the way.   The process according to claim 7, wherein the solvent used in step (a) is selected from hexane, n-pentane or cyclohexane.   8. The method of claim 7, wherein the yield of pipetalin is about 1.2 weight percent based on the dried fruit.   8. The method of claim 7, wherein the sesamin yield is about 0.04 weight percent based on the dried fruit.   8. The method of claim 7, wherein the yield of peritrine is about 0.04 weight percent based on the dried fruit.   8. The method of claim 7, wherein the yield of guaneancin is about 0.06 weight percent based on the dried fruit.   8. The method of claim 7, wherein the yield of brachystamide-B is about 0.024 weight percent based on the dried fruit.   8. The method of claim 7, wherein the purity of the compound obtained from the above method is up to 90%.

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