Chemical composition and anticholinesterase activity of lagochilus inebrians D. Kh. Akramov,1

DOI 10.1007/s10600-019-02748-x
Chemistry of Natural Compounds, Vol. 55, No. 3, May, 2019
CHEMICAL COMPOSITION AND ANTICHOLINESTERASE
ACTIVITY OF Lagochilus inebrians
D. Kh. Akramov,¹ M. Bacher,² G. Zengin,³ S. Bohmdorfer,² T. Rosenau,² Sh. S. Azimova,¹ and N. Z. Mamadalieva^1*
The plant genus Lagochilus (Lamiaceae) comprises 44 species that are broadly distributed over Central Asia with 18
species occurring in the flora of Uzbekistan [1]. Tincture of the herb and roots of L. inebrians Bunge, which is indigenous in Uzbekistan [2], are used in folk medicine as a sedative, analgesic, and hemostatic agent [3]. The hemostatic activity is due to the diterpene alcohol lagochilin. Hemostatic materials based on this compound were fabricated [4]. Diterpenes (lagochilin and its acetates) [5–7] and iridoids (8-O-acetylharpagide and harpagide) [8] were observed in L. inebrians. However, the chemical composition of this plant is inadequately studied. The enzyme-inhibiting activity has not been studied.
The BuOH extract of the aerial part of L. inebrians afforded 5-hydroxy-7,4′-dimethoxyflavone (1), β-sitosterol (2) ,
stigmasterol (3), daucosterol (4), lagochilin (5) [5], and 8-O-acetylharpagide (6) [8]. Compounds 1–4 were isolated for the first time from this plant. The chemical structures were identified using IR, mass, and NMR spectra that were taken on the instruments described earlier [9, 10].
Aerial parts of L. inebrians were collected on June 26, 2017, in Ukhum Pass in the Nuratau mountains of Jizzakh Oblast. Taxonomic identification came from the Institute of Botany, Academy of Sciences of Uzbekistan. A specimen of L. inebrians is preserved in the herbarium collection under code No. 1768. The species was determined by Dr. A. R. Batoshev.
Milled air-dried aerial parts of L. inebrians (2 kg, <5% moisture) were extracted with MeOH (5 × 8 L). The resulting MeOH extracts were combined and evaporated at 40°C. The condensed extract (160 g, 8% calculated for air-dried raw material) was dissolved in distilled H₂O (1:1) and fractionated sequentially by polarity using CHCl₃ (4 × 500 mL) to remove low-molecular-mass slightly polar compounds and n-BuOH (4 × 500 mL). Then, the BuOH extract was evaporated to afford dry extract (75 g, 3.75%). The BuOH extract was mixed with silica gel and placed onto a chromatography column of silica gel (100–200 mesh, Tianjin Sinomed Pharmaceutical, China) in a 30:1 ratio to the amount being separated. Compounds were eluted by solvent systems CHCl₃ (fraction A); CHCl₃–MeOH (15:1, B; 9:1, C; 4:1, D); and CHCl₃–MeOH–H₂O (4:1:0.1, E). Recrystallization of fraction A (2.3 g) produced 1 (20 mg). Fraction B (10.2 g) was rechromatographed over a column of silica gel with elution by hexane–Me₂CO (25:1) to isolate 2 (13 mg) and subfraction BA (4.3 g). Subfraction BA was chromatographed over silica gel (CHCl₃ and CHCl₃–MeOH, 50:1) to separate 3–5 (8, 7, and 128 mg, respectively). Fraction D (9.6 g) was separated over silica gel using CHCl₃–MeOH (9:1) to isolate 6 (54 mg).
Lagochilus (Lamiaceae) o'simlik jinsi 44 turni o'z ichiga oladi, ular O'rta Osiyoda 18 tasi bilan keng tarqalgan.
Oʻzbekiston florasida uchraydigan turlar [1]. Oʻzbekistonda mahalliy boʻlgan L. inebrians Bunge oʻti va ildizining damlamasi [2] xalq tabobatida tinchlantiruvchi, ogʻriq qoldiruvchi, qon toʻxtatuvchi vosita sifatida qoʻllaniladi [3]. Gemostatik faollik lagoxilin diterpen spirtiga bog'liq. Ushbu birikma asosida gemostatik materiallar ishlab chiqarilgan [4]. L. inebrianlarda diterpenlar (lagoxilin va uning asetatlari) [5-7] va iridoidlar (8-O-asetilharpagid va harpagid) [8] kuzatilgan. Biroq, bu o'simlikning kimyoviy tarkibi etarli darajada o'rganilmagan. Fermentni inhibe qiluvchi faolligi o'rganilmagan.
L. inebriansning havo qismining BuOH ekstrakti 5-gidroksi-7,4'-dimetoksiflavon (1), b-sitosterol (2) ni,
stigmasterol (3), daukosterol (4), lagochilin (5) [5] va 8-O-asetilharpagid (6) [8]. 1-4 birikmalar birinchi marta bu o'simlikdan ajratilgan. Kimyoviy tuzilmalar ilgari tasvirlangan asboblarda olingan IQ, massa va NMR spektrlari yordamida aniqlandi [9, 10].
L. inebriansning havo qismlari 2017-yil 26-iyun kuni Jizzax viloyatining Nurotau tog‘lari hududidagi Uxum dovonida yig‘ilgan. Taksonomik identifikatsiya O‘zbekiston Fanlar akademiyasi Botanika institutidan olingan. L. inebrians namunasi gerbariy kolleksiyasida 1768-raqamli kod boʻyicha saqlanadi. Turni doktor A. R. Batoshev aniqlagan.
L. inebrians ning maydalangan havoda quritilgan havo qismlari (2 kg, <5% namlik) MeOH (5 × 8 L) bilan ekstraksiya qilingan. Olingan MeOH ekstraktlari birlashtirildi va 40 ° C da bug'lanadi. Quyultirilgan ekstrakt (160 g, havoda quritilgan xom ashyo uchun hisoblangan 8%) distillangan H2O (1: 1) da eritildi va past molekulyar massali ozgina qutbli birikmalarni olib tashlash uchun CHCl3 (4 × 500 ml) yordamida polarit bo'yicha ketma-ket fraktsiyalarga bo'lindi. va n-BuOH (4 × 500 ml). Keyin BuOH ekstrakti bug'lanib, quruq ekstrakt (75 g, 3,75%) hosil bo'ldi. BuOH ekstrakti silika jeli bilan aralashtirildi va ajratilayotgan miqdorga 30:1 nisbatda silikagelning xromatografiya ustuniga (100-200 mesh, Tianjin Sinomed Pharmaceutical, Xitoy) joylashtirildi. Murakkablar CHCl3 (A fraktsiyasi) erituvchi tizimlari bilan elutildi; CHCl3-MeOH (15:1, B; 9:1, C; 4:1, D); va CHCl3-MeOH-H2O (4:1:0,1, E). A fraktsiyasining qayta kristallanishi (2,3 g) 1 (20 mg) hosil qildi. B fraksiyasi (10,2 g) 2 (13 mg) va BA subfraktsiyasini (4,3 g) ajratib olish uchun geksan-Me2CO (25:1) bilan elyusiya bilan silikagel ustuni ustida rexromatografdan o'tkazildi. BA subfraktsiyasi 3-5 (mos ravishda 8, 7 va 128 mg) ni ajratish uchun silika jeli (CHCl3 va CHCl3-MeOH, 50:1) ustida xromatografiya qilindi. D fraksiyasi (9,6 g) 6 (54 mg) ni ajratib olish uchun CHCl3-MeOH (9:1) yordamida silikagel ustida ajratildi.