JP2008150309A - Brandisianin a, b and c, and medicament using the same - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a new chemical substance as a resource. <P>SOLUTION: The brandisianin A is represented by the formula. The invention is utilizable in industry as a medicament containing the new compound. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to brandicyanines A, B and C, which are novel compounds, and further to a drug using them.

  In our current life, chemicals contained in the body of natural animals and plants, microorganisms, etc. (hereinafter referred to as “natural products”) are those that have a beneficial effect on the human body and are effective in crude drugs and pharmaceuticals. Used as an ingredient. Moreover, such a thing has various roles also as a research material for developing a more useful pharmaceutical, and is very important.

  As described above, as a report on the search for natural products that have a beneficial effect on the human body, for example, Non-Patent Document 1 listed below extracts a bisindole compound, a naphthoquinone compound, a glyceride compound and the like from a deformed bacterium.

Masami Ishibashi, “Recycling unused fungi: Search for natural products from deformed bacteria”, Journal of Synthetic Organic Chemistry, 2003, Vol. 61, No. 2, pp. 152-163

  However, on the other hand, despite the fact that many people have been searching for natural products, less than 10% of all the species on Earth have been studied and investigated as search materials. It is said.

  An object of this invention is to provide a novel chemical substance as a resource.

The chemical substance according to one means of the present invention is represented by the following formula (1). The chemical substance represented by the following formula (1) is called brandicyanin A.

Moreover, the chemical substance which concerns on another means of this invention is shown by following formula (2). The chemical substance represented by the following formula (2) is called brandicyanin B.

A chemical substance according to another means of the present invention is represented by the following formula (3). The chemical substance represented by the following formula (3) is called brandicyanin C.

Moreover, the chemical | medical agent which concerns on another means of this invention contains the brandicyanine A shown by following formula (1) as an active ingredient.

In addition, a cancer drug according to another means of the present invention contains brandicyanin B represented by the following formula (2) as an active ingredient.

Moreover, the chemical | medical agent which concerns on another means of this invention contains the brandicyanine C shown by following formula (3) as an active ingredient.

  In addition, although said medicine is not necessarily limited, it is anticipated that it will be useful as a cancer therapeutic agent.

  According to the present invention, a novel chemical substance can be provided as a resource. In particular, the chemical substance according to the present invention is expected to be used as, for example, a therapeutic agent for cancer because it exhibits a cell growth inhibitory effect on cancer cells.

  Hereinafter, embodiments of the present invention will be described.

Brandicyanines A, B, and C according to an embodiment of the present invention are represented by the following formulas (1) to (3), respectively.

  Brandicyanines A, B, and C according to the present embodiment can be extracted from the leguminous plant Millettia brandisiana as is clear from the examples described later, but the invention is not limited thereto, and synthesis is also considered possible. It is done.

  In addition, the brandicyanins A, B, and C according to this embodiment are expected to be used as drugs, for example, cancer therapeutics, because they exhibit a cell growth inhibitory action against cancer cells. In the case where brandicyanins A, B and C are used as cancer therapeutic agents, it is preferable to contain at least one of brandicyanines A, B and C and salts thereof as active ingredients.

  In addition, the therapeutic agent for cancer according to the present embodiment includes at least one of the above-mentioned brandicyanins A, B, and C and salts thereof, as well as usual pharmaceutically acceptable carriers, binders, stabilizers, enhancers. It may contain various preparation ingredients such as a form, a diluent (eg, distilled water), a pH buffer (eg, phosphate buffered saline), a disintegrant, a solubilizer, a solubilizer, and an isotonic agent. it can.

  Moreover, the therapeutic agent for this cancer can be administered orally or parenterally in an appropriate dose suitable for the sex, weight and symptoms of the patient. For oral administration, commonly used administration forms such as powders, granules, tablets, capsules, solutions, suspensions, oils, emulsifiers and the like can be employed. In addition, as parenteral administration, a commonly used administration form, for example, a method of administering the above-mentioned solution, suspension or the like directly to the affected area, a form of administration by injection or the like can be adopted.

  This example shows the results of examination of blandicyanin A extracted from leguminous plant Millettia brandisiana. FIG. 1 shows an outline of the scheme for the isolation of bradycyanins A, B and C.

  First, 30.5 g of a methanol extract of the leaf part of Millettia brandisiana was obtained, applied to a column (φ43 × 400 mm) using Diaion HP-20 as a carrier, and 1A (23. 66 g), 1B (0.36 g) and 1C (3.18 g).

  Next, about 1A, solvent distribution was performed using n-hexane, ethyl acetate, and butanol, and each soluble part was obtained. Among these, the ethyl acetate-soluble part was attached to a column (φ30 × 350 mm) using Silica gel PSQ 100B as a carrier, eluted with n-hexane, acetone and methanol, and 2A (2.4 g) in the order of elution. Each fraction of 2B (0.32 g) and 2C (1.7 g) was obtained.

  Further, 2A was subjected to preparative HPCL (ODS) under the HPLC conditions (1) described in Table 1 below to obtain brandicyanin A (13.1 mg) and brandicyanin B (23.4 mg).

Further, 2B was applied to a column (φ1.6 × 500 mm) using Silica gel 60N as a carrier, eluted with n-hexane, acetone and methanol, and separated into 7 fractions of 3A to 3G in the order of elution. . And about 3A, it attached | subjected to preparative HPLC (ODS) by the HPLC conditions (2) shown in following Table 1, and obtained the brandicyanine C (10.0 mg).

(Brandicyanine A)
Next, high-resolution FABMS was performed on the brandicyanin A, and a peak at m / z 437.1004 presumed to be [M + K] ⁺ was observed. As a result, the molecular formula was estimated as C ₂₂ H ₂₂ O ₇ .

Next, infrared absorption measurement was performed to obtain an infrared absorption spectrum (hereinafter referred to as “IR spectrum”). As a result, absorptions considered to be derived from OH groups and carbonyl groups were observed at 3263 cm ⁻¹ and 1658 cm ⁻¹ , respectively.

Further, ultraviolet absorption measurement was performed on brandicyanin A, and an ultraviolet absorption spectrum (hereinafter referred to as “UV spectrum”) was also obtained. This ultraviolet absorption measurement was performed using methanol as a solvent with a concentration of 2.0 × 10 ⁻⁵ mol / l and a cell length of 1 cm. As a result, it has absorption peaks at 295.5 nm, 260.0 nm, and 203.5 nm, respectively, and it was estimated that the compound was an aromatic compound or a conjugated double bond compound. The molar extinction coefficient (ε) in each of them was 11550, 21250, and 37950, respectively. FIG. 2 shows the UV spectrum of brandicyanin A.

  Further, measurement of electron impact mass spectrometry (EIMS) was performed on brandicyanin A. As a result, it was found that the molecular weight of this compound was 398.

In addition, the optical rotation angle [α] _D was measured for brandicyanin A. As a result, the optical rotation angle was 0 degree, and it was confirmed that it was optically inactive. Table 2 below shows high-resolution FABMS, EIMS, IR spectrum, UV spectrum, and optical rotation angle data for brandicyanin A.

Furthermore, ¹ H NMR and ¹³ C NMR were also measured for brandicyanin A. The results are shown in Table 3, FIG. 3 and FIG. As a result, in this substance, signals derived from the isoflavone skeleton and signals derived from a series of prenyl groups were observed. Further, the signal of the carbonyl carbon δc180.8 found signal from hydroxyl groups which are estimated to bind the carbonyl carbon and hydrogen [delta] _H 12.91 was observed.

  Furthermore, HMQC and HMBC spectrum analysis was also performed on the brandicyanin A. FIG. 5 shows an HMQC spectrum, and FIG. 6 shows an HMBC spectrum. As a result, a HMBC correlation was observed from the 1 ″ -position methylene proton to the 4′-position of the prenyl group, and from the 1 ″ -position chemical shift value, the prenyl group was bonded to the 4′-position of the B ring via an oxygen atom. It was recognized that

From the above, the structure of brandicyanin A was determined as the following formula.

Further, when the IC ₅₀ value for HeLa cells was measured for this brandicyanin A, it was confirmed that the value was larger than 25 μg / ml, and at this concentration, the growth inhibitory action against cancer cells could not be confirmed. It was.

(Brandicyanine B)
Next, high resolution FABMS was performed on the brandicyanin B, and a peak at m / z 396.1212 estimated to be [M ⁺ ] was observed. As a result, the molecular formula was estimated as C ₂₂ H ₂₀ O ₇ .

Next, infrared absorption measurement was performed to obtain an IR spectrum. As a result, the absorption considered to be derived from the OH group and the carbonyl group was observed at 3545 cm ⁻¹ and 1658 cm ⁻¹ , respectively.

Further, UV absorption measurement was performed on brandicyanin B, and a UV spectrum was also obtained. This ultraviolet absorption measurement was performed using methanol as a solvent with a concentration of 2.0 × 10 ⁻⁵ mol / l and a cell length of 1 cm. As a result, it has absorption peaks at 294.5 nm, 263.5 nm, and 202.5 nm, respectively, and could be estimated to be an aromatic compound or a conjugated double bond compound. The molar extinction coefficient (ε) in each of them was 10200, 24500, and 40950, respectively. FIG. 7 shows the UV spectrum of brandicyanin B.

  Further, measurement of electron impact mass spectrometry (EIMS) was performed on brandicyanin B. As a result, it was found that the molecular weight of this compound was 396.

In addition, the optical rotation angle [α] _D was measured for brandicyanin B. As a result, the optical rotation angle was −4.77 degrees, confirming optical activity. Table 4 below shows high-resolution FABMS, EIMS, IR spectrum, UV spectrum, and optical rotation angle data for brandicyanin B.

Furthermore, ¹ H NMR and ¹³ C NMR were also measured for brandicyanin B. The results are shown in Table 5, FIG. 8 and FIG. This suggested the presence of an isopropenyl dehydrofuran ring.

  Furthermore, HMQC and HMBC spectrum analysis was also performed on the brandicyanin B. FIG. 10 shows an HMQC spectrum, and FIG. 11 shows an HMBC spectrum. As a result, a HMBC correlation was observed from the methylene proton at the 1 ″ position of the prenyl group to the 4 ′ position, and the prenyl group was bonded to the 4 ′ position of the B ring via an oxygen atom from the chemical shift value at the 1 ″ position. It was recognized that

From the above, the structure of brandicyanin B was determined as the following formula.

Further, when the IC ₅₀ value for HeLa cells was measured for this brandicyanin B, it was confirmed to be 9.7 μg / ml, and the growth inhibitory action against cancer cells could be confirmed.

(Brandicyanine C)
High resolution FABMS was performed on the brandicyanin C, and a peak at m / z 398.1370 estimated to be [M ⁺ ] was observed. As a result, the molecular formula was estimated as C ₂₂ H ₂₂ O ₇ .

Next, infrared absorption measurement was performed to obtain an IR spectrum. As a result, absorption considered to be derived from the OH group and the carbonyl group was observed at 3469 cm ⁻¹ and 1655 cm ⁻¹ , respectively.

Further, UV absorption measurement was performed on brandicyanin C, and a UV spectrum was also obtained. This ultraviolet absorption measurement was performed using methanol as a solvent with a concentration of 2.0 × 10 ⁻⁵ mol / l and a cell length of 1 cm. As a result, it had absorption peaks at 293.0 nm, 264.0 nm, and 202.5 nm, respectively, and could be estimated to be an aromatic compound or a conjugated double bond compound. The molar extinction coefficient (ε) in each of them was 8250, 18000, and 31450, respectively. FIG. 12 shows the UV spectrum of brandicyanin A.

  Further, measurement of electron impact mass spectrometry (EIMS) was performed on brandicyanin C. As a result, it was found that the molecular weight of this compound was 398.

Further, the rotation angle [α] _D was measured for the brandicyanin C. As a result, the optical rotation angle was 0 degree, and it was confirmed that it was optically inactive. Table 6 below shows high resolution FABMS, EIMS, IR spectrum, UV spectrum, and optical rotation angle data for brandicyanin A.

  Furthermore, 1H NMR and 13C NMR were also measured for brandicyanin C. The results are shown in Table 7, FIG. 13 and FIG. As a result, a signal derived from the isoflavone skeleton and a signal derived from the prenyl group were observed in this substance.

  Furthermore, HMQC and HMBC spectrum analysis was also performed on the brandicyanin C. FIG. 15 shows an HMQC spectrum, and FIG. 16 shows an HMBC spectrum. As a result, a correlation from the methylene proton at the 1 ″ position of the prenyl group to the carbons at the 7, 8, and 8a positions was observed, and it was confirmed that the prenyl group was bonded to the 8-position of the A ring.

From the above, the structure of brandicyanin C was determined as the following formula.

Further, when the IC ₅₀ value for HeLa cells was measured for this brandicyanin C, it was confirmed to be 19.1 μg / ml, and the growth inhibitory action against cancer cells could be confirmed.

  The present invention has industrial applicability as a drug containing a novel compound.

FIG. 2 is a diagram showing an outline of a scheme for isolation of bradycyanin A, B and C. It is a figure which shows the UV spectrum of brandycyanin A. It is a diagram showing the ¹ H NMR spectrum of Branzi cyanine A. It is a figure which shows the ¹³ C NMR spectrum of brandy cyanine A. It is a figure which shows the HMQC spectrum of brandycyanin A. It is a figure which shows the HMBC spectrum of brandy cyanine A. It is a figure which shows the UV spectrum of brandycyanin B. It is a diagram showing the ¹ H NMR spectrum of Branzi cyanine B. It is a figure which shows the ¹³ C NMR spectrum of brandy cyanine B. It is a figure which shows the HMQC spectrum of brandy cyanine B. It is a figure which shows the HMBC spectrum of brandy cyanine B. It is a figure which shows the UV spectrum of brandycyanin C. It is a diagram showing the ¹ H NMR spectrum of Branzi cyanine C. It is a figure which shows the ¹³ C NMR spectrum of brandycyanin C. It is a figure which shows the HMQC spectrum of brandycyanin C. It is a figure which shows the HMBC spectrum of brandy cyanine C.

Claims (7)

Bradycyanin A represented by the following formula.
Bradycyanin B represented by the following formula.
Bradycyanin C represented by the following formula.
A drug containing bradycyanin A represented by the following formula or a salt thereof as an active ingredient.
A drug containing bradycyanin B represented by the following formula or a salt thereof as an active ingredient.
A drug containing bradycyanin C represented by the following formula or a salt thereof as an active ingredient.
  The drug according to claim 4, 5 or 6, which is a therapeutic drug for cancer.