DE60216986T2 - NEW USE OF PIPATALIN - Google Patents

Description

The Invention describes a method for α-glucosidase inhibition in a Subject by administering a pharmaceutical composition, the pipataline (Formula 1a). In particular, the invention relates the isolation of pipataline [5-1-dodecenyl) -1,3-benzodioxole] the plant source Piper longum in significant yields. The invention further identifies the therapeutic application of this compound as an α-glucosidase inhibitor in the form of a suitable pharmaceutical composition for treatment of diseases such as hyperlipoproteinemia, viral infection, heptitis B and C and HIV.

State of technology

The Use of plants as medicine extends to early humanity back. In fact, the high cultures of the ancient Indians, Chinese supplied and North Africans written evidence of the wealth of sentiment Humanity in the use of plants to treat numerous Diseases (Phillipson J.D., Phytochemistry, 2001, 56, 237-243). With the extension of findings through new research results and clinical experiences are also changes in drug therapy to observe. As a result of the emergence of new diseases and the Identification of targets with multiple therapeutic applications Hold the Search for new compounds with unique structures and properties continue on.

The α-glucosidase enzyme was identified as such a target. α-Glucosidasehemmer find increasingly therapeutic Use in metabolic diseases such as diabetes mellitus, obesity, hyperlipoproteinemia Type IV (Truscheit, E. et al., Angew Chem. Int. Ed. Engl., 1981, 20, 744-761; Puls, W., Keupu Diabelologia, 1973, 9, 97, Pulse, W., Habilitation thesis University Dusseldorf, 1980), HIV, human hepatitis B virus, human cytomegalovirus and Influenza (Heightman, T.D. and Vasella, A.T., Angew. Chem. Int. Ed. Engl., 1999, 38, 750-770; Mehta et al. FEBS Lett., 1998, 430, 17-22; Watson, A.A. et al., Phytochemistry, 2001, 56, 265-295), Cancer and in immunocompromised cases. It was found that the serum levels of glucosidases in many patients with different Tumors increased (Wollen, J.W. and Tesiar, P., 1965, Din. Chem. Ada., 12, 671-683), and it was recognized that they are involved in tumor cell invasion at degradation the extracellular Matrix are involved (Bernaki, R.J. et al., 1985, Cancer Metastasis Rev., 4, 81-102). Therefore, one goes to inhibitors of catabolic glucosidases active for a therapeutic strategy against cancer (Watson, A.A. et al., Phytochemistry, 2001, 56, 265-295).

A Variety of compounds with potential as α-glucosidase inhibitors has been on their chemotherapeutic value (EI Ashry et al., Pharmacology, 2000, 55, 251-262, 331-348 and 403-415). Although several active ingredients for α-glucosidase inhibition either used clinically or in different stages clinical development (Drugs of the future, 1986, 11, 795-797; Drugs of the future, 1986, 11, 1039-1042; Watson, A. A. et al., Phytochemistry, 2001, 56, 265-295), emphasizes the Impact of the disease burden discussed above obvious Need for new resources. Furthermore is a big Pool of inhibitors necessary as patients against the current Medications can develop resistance.

Historical have the knowledge gained through traditional medicine and that Screening of extracts of plants with animals, new natural Products supplied by themselves are potential bioactive agents for Treatment of human diseases (Gullo, V. P., The discovery of natural products with therapeutic potential, Butterworth-Heinemann, Boston, 1994; Cragg, G., Metal J. Nat. Prod., 1997, 60: 52-60).

The Screening natural Sources led to discover many clinically useful Agents not only in the treatment of those discussed above Diseases, but also in the prevention of such diseases a big Role-play. The increasing clinical significance of epidemics diabetes, cancer, HIV and other viral diseases as well as Drug resistance makes identifying new sources effective Connections to maintain a large pool therefore even more urgent.

As described below led our search for α-glucosidase inhibitors from traditional medicinal plants to identify piper longum, the effective α-glucosidase inhibitor in big Amount contained.

Piper longum Linn. (Pippali) has been described in traditional Indian medicine for malaria fever, heart disease, splenomegaly, whooping cough, edema, etc. (PV Sharma, Classical uses of medicinal plants, Haridas Ayurvedic series (4), Chaukambha Viswabharathi, Varanasi, 1996).

The The present invention relates to identification and isolation of pipataline (1a) as an effective α-glucosidase inhibitor from Piper longum in the form of a suitable pharmaceutical composition, in the treatment of diseases such as hyperlipoproteinemia, viral infection, Hepatitis B and C and HIV can be used.

Various Species of pepper (Piper) and claimed according to the invention as α-Glucosidaseinhibitoren Compounds thereof are listed in Table 1 and their biological activity is in Table 2 given.

Application and administration:

The α-glucosidase inhibitor of this invention may be used according to any conventional method of management and for the treatment of HIV, AIDS, hepatitis B and / or C and others Viral infections are applied or administered.

to Application in humans and animals and / or in veterinary medicine Applications may be the α-glucosidase inhibitors according to the invention according to convenience and clinical condition be administered in different ways. For use in humans can the α-glucosidase inhibitors accordingly the requirement of the case over different routes, including oral, intraperitoneal, intravenous and / or intramuscular Ways.

formulations:

Of the α-Glucosidasehemmer according to the invention can be combined with any pharmaceutically acceptable additive, carrier and Formulated vehicle, the effectiveness and property should not change the connection in any case.

For applications In humans, the α-glucosidase inhibitor according to the invention can be used in a person skilled in the art known manner with numerous pharmaceutically acceptable carriers and additives that are prescribed for administration of the pharmaceutical are suitable.

The chosen carrier or vehicle should be natural with the application or Administration of glucosidase inhibitors.

Effective amounts:

The Terms "an effective amount" and / or "an inhibitory amount" refer to the amount on α-glucosidase inhibitor compound according to the invention, which seems necessary in comparison with an untreated one Control under suitable treatment conditions depends on Disease condition and severity of the disease a reduction of the disease Disease severity, such as significant suppression of viral infection receive. This also means that an effective amount of the α-glucosidase inhibitor compound The invention should be less than an amount due to a particular disease-treated organism has significant undesirable side effects causes. This implication is supported by the use of the term "pharmaceutically effective Quantity "affirmed actual Application rate and amount may vary depending on the condition vary. This may be the case regardless of the concentrations as described in the examples of the invention.

The actual Application rate and amount may vary depending on the severity of the disease and / or the infection may vary and may also depend on a variety of factors such as the age and sex of the individual treated and the mode of administration etc., depend. Considering These factors should be the actual ones Dose level and dosage regimen by one of ordinary skill in the art can be determined without further ado.

Table 1:

pepper connections show numerous different biological activities. The biological activity of Pipataline, sesamin, pellitorin, guineensin and brachystamide-B in Table 2.

Table 2:

The The main object of the invention is that obtained from Piper longum Pipataline has a new effect as an α-glucosidase inhibitor provide.

isoliert aus Piper longum, und eines pharmazeutisch verträglichen Zusatzstoffs zur Herstellung einer pharmazeutischen Zusammensetzung zum Management und zur Behandlung von Krankheiten wie Hyperlipoproteinämie, Virusinfektion, Hepatitis B und C und HIV.Accordingly, the invention relates to the use of an effective amount of pipataline of formula 1a:

isolated from Piper longum, and a pharmaceutically acceptable additive for the manufacture of a pharmaceutical composition for the management and treatment of diseases such as hyperlipoproteinemia, viral infection, hepatitis B and C and HIV.

The The present invention relates to a new effect of Piper longum received pipatalin as an α-glucosidase inhibitor in the management and treatment of human diseases, like hyperlipoproteinemia, Viral infection, hepatitis B and C and HIV.

The The invention further relates to the isolation of pipaline from a new source, namely Piper longum.

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

The The present invention provides, for the first time, a process for isolation of pipaline from Piper longum ready.

• a. Extraktion der getrockneten Piper longum-Früchte mit einem Lösungsmittel,
• b. Einengen des Extrakts unter Vakuum unter Erhalt eines Rückstands, und
• c. Eluieren des Rückstands von Schritt (b) mit Hexan unter Erhalt von Pipatalin und einem Rückstand.

The process for isolation of pipataline (Formula 1a) comprises the steps:

• a. Extraction of the dried Piper longum fruits with a solvent,
• b. Concentrate the extract under vacuum to give a residue, and
• c. Elute the residue of step (b) with hexane to give pipataline and a residue.

at an embodiment For example, the solvent used in step (a) is selected from Hexane, cyclohexane or n-pentane.

According to the invention the isolation of pipataline from a completely new source.

Short description of Drawings:

The Invention is illustrated by the attached drawings, in which:

1 (a) the formula of pipataline is [5- (1-dodecenyl) -1,3-benzodioxole];

2 (a) Figure 4 is a graph showing the α-glucosidase inhibitory activity of pipataline, sesamin, pellitorin, guineensin and brachystamide-B.

Some the embodiments of the present invention are illustrated in the following examples, not as restrictions the scope of the invention should be understood.

example 1

Experimental procedure for isolating pipataline.

The Dried, powdered Piper longum fruits (500 g) were grown on a Soxhlet apparatus loaded. The powder was extracted with hexane. The hexane extract was concentrated in vacuo. The dark green colored residue was up to an altitude 60 cm loaded silica gel column loaded with 60-120 mesh and 3.5 cm diameter.

To the Receipt of pipataline became the pillar initially eluted with hexane. The pipataline yield is about 6.0 g.

The the above compound was obtained in a purity of 90%.

The spectrochemical and physical properties of the above compound are as follows:
Pipataline 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.
¹ H-NMR (200 MHz, CDCl ₃ ) (δ)
0.95 (3H, t, H-1), 1.20-1.60 (16H, b, H-2-9), 2.20 (2H, q, H-10), 5.90 (2H , s, -OCH ₂ O), 6.0-6.15 (1H, q, H-11), 6.30 (1H, d, J = 15.5 Hz, H-12), 6.70 ( 2H, s, H-5 ', 6'), 6.88 (1H, s, H-2 ¹ ).
¹³ C-NMR (50 MHz, _CDCl3)
14.12 (C-1), 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- ^6f ), 129.27 (C-11), 129.57 (C -12), 132.61 (C-1 '), 146.68 (C-4'), 148.01 (C-3 ').
EI-MS: M ⁺ 288

Example 2:

Determination of α-glucosidase inhibiting Effect of P. longum isolated compound:

Of the Test for α-glucosidase inhibition was carried out by a chromogenic method. In short, were 10 μl test compound, solved in DMSO (5 mg / ml and further dilutions), 5 min with 5 ul of α-glucosidase enzyme from yeast in 100 mM phosphate buffer (pH 7.00). After 5 minutes incubation 50 μl of 5 mM substrate (p-nitrophenyl-α-D-glucopyranoside in the same Buffer) was added. The absorbance at 405 nm before substrate addition and 5 min after substrate addition was recorded spectrophotometrically. The increase in extinction from before substrate addition to reaction after substrate addition was obtained. The percentage inhibition was calculated according to (1 - OD test / OD control) × 100, and the 50% inhibitory concentration (IC50) was determined by appropriate Regression analysis calculated.

According to the invention was found that pipatalin is isolated from Piper longum, which is a completely new Source is. The yield of this compound is also substantial. Further, it has been found that the above compound has the property α-glucosidase inhibition shows.

Advantages:

α-glucosidase have youngest because of their wide spectrum of activity in diseases different Origin such as viral diseases, HIV, hepatitis B and C etc. attention obtained. Much attention is now being devoted to the a-glucosidase inhibitors from natural Provide sources.

The Compound pipataline is used in pure form. The insulation of pipaline from Piper longum in significant yields as an α-glucosidase inhibitor makes the invention therefore very significant.

Claims (3)

Use of an effective amount of pipataline of the formula 1a:

isolated from Piper longum, and a pharmaceutically acceptable additive for the manufacture of a pharmaceutical composition for the management and treatment of diseases such as hyperlipoproteinemia, viral infection, hepatitis B and C and HIV. Use according to claim 1, wherein pipataline is an α-glucosidase inhibiting Has an effect of up to 77.45%. Use according to claim 1 or 2, wherein pipataline an IC50 value of 26.52 μg / ml Has.