Isolation or ecdysterone (crustecdysone) from polypodium vulgare L. Rhizomes

TetrahedronLetter8No.18, pp. 1689-1691, 1967.

PergamonReae Ltd. Printedin GreatBritain.

ISOLATION OF ECD~STNR~NN (CRHSTECBYS~NE) mot4 POLYPODIUM WLOARE

L. RHIZ~NES

J. Jisba, V. Berout and F. sorm Institute of Organic Chemistry and Biochemistry, Ceechoslovak Academy of Sciences, Prague [Received15 February 1967)

As a part of study (1) of constituents of p0iyp0ay rhizomes(Polynodium vulgare L.) we isolated recently, besides other compounds, also two crystalline substances and proposed the names polypodine A and polypodine B for them.*

We have found now (3) polypodine A to be identical with ecdysterone

(hydroxyecdysone, crustecdysone) which, as has been shqwn, is the moulting hormone isolated from Crustaceans (4) and later from Bombyx mori (5)

and has

the structure of 2p,3B,14a,205,22BF,25-hexahydroxy-5B-cholest-7-en-6-one ; ecdysterone possesses higher physiological activity in comparison with ecdysone isolated from insects.

The proof of identity of polypodine A with

ecdysterone has been carried out by comparison of physical constants of both compounds, chemical properties and biological activity.

The physical constants

are summarized in Table I. Ecdysterone crystallizes from an aqueous solution with three molecules of water (like ecdysone which crystallizes with two molecules of water) and as a hydrate has m. p. 150 - 151'C. analysis corresponds to the molecular 60.65% C, 9.43% H; found

The elemental

formula C 27H4407'3H20 (534.7) calculated

60.28% C, 9.23% H.

The molecular weight has been determined by mass spectrometry (M = 4801; this is in accordance with the molecular weight of tetra(trimethylsilylether) prepared from polypodine A (hi= 768) as well. The character of fragments in the mass spectra was entirely ic?enticalin both compounds and the NMR spectra as well are in full accordance.

The i,nfra-

*A substance called polypodine (m. p; 150 - 151'C) was, for the first time, reported by Volmar and Reebe (2). The authors took polypodine for a glycoside and did not characterize it in detail.

16139

1690

No.18

TABLE Physical

Constants

of

I

Polypodine

A and Ecdysterone

Polypodine p.

m.

A

Ecdysterone

243’C

237.5

+61.8’C

MB W

(methanol)

- 239.5Oc

l

244 nm (logr

4.09)

242 nm (log

& 4.09

240 nm (logg IR (KBr disc)

*For red

ecdysone spectra

of

maxima of double

the

were

found

la],,

our

specimen,

identical observed

temperatures,

the

ecdysterone.

This

1608

cm-l

(6)

1650,

1610

cm-l

(5)

1645,

1612

cm-’

(6)

at

80°C),

+58.5’C measured

main frequencies,

bond,

there

1642,

(5)

4.13)

with

particularly the

certain

in KBr pelet of

published

infra-red

spectrum

fenomenon

is

carbonyl

data.

differences.

group

In other

After

drying

was entirely

quite

(dried

frequent

identical and has

and conjugated

parts

the

exhibited

of

the

spectra

sample

at

higher

with

that

(6)

been

observed

of

already

many times. Polypodine sulfate For

A on treatment

yielded

C33H5207

70.45%

C,

presence

a characteristic (560.8)

9.44% of

potassium

periodate

does

has been

All

take

the

The presence in

tested

place

PolyoOdiUn

of

the

groups,

prove

high

rhizomesindicates

l-I+;

229-234’C.

found

:

corroborates

as well

activity

larvae

50 yg]

of

as the

days

in

the

result

insect.

Pyrrbocoris

implanted

two and a half

the

H, 0.36%

cupric

of

polarographically).

moulted

seventh

m. p.

diac,etonide

ecdysone-like

(about

after

a considerably

vulgare

hydroxyl

of anhydrous

derivative,

of

on freshly

discussed

presence

The formation

(determined

compound

the

C, 9.35%

vicinal

before

in

: 70.68%

enormous

moulting

facts

H+.

oxidation

of the

precocious not

of

A exhibits

microcrystals cause

H, 0.38%

acetone

diisopropylidine

calculated

two pairs

Polypodine effect

with

in

to

whereas

The The

apterus.

the

haemolymph

the

normal

moulting

day. identity

amount that

of of

polypodine

ecdysterone compounds

A with (over

of

the

ecdysterone.

1% of character

dry of

drug’

1691

No.18

moulting hormone may be exogenous factors and that the insect receives them with the food.

Indirectly this assumption is in agreement with the fact that

compounds of ecdysone activity were isolated from adult insect specimens, in which the phoracic glands had already degenerated. REFERENCES 1.

J. Jizba and V. Herout, Coll. Czech. Chem. Commun., in press.

2.

J. Volmar and E. Reebe, J. Am. Pharm. Assoc., 22, 1225 (1933).

3.

J. Jizba, V. Herout and F. Sorm, Czechoslovak patent application.

4. 5.

F. Hampshire and D.H.S. Horn, Chem. Comm.,1966, ------- 37. P. Hocks and R. Wiechert, Tetrahedron Letters, i.$g, 2989.

6.

H. Hoffmeister and H.F. 'riitzmacher,Tetrahedron Letters, 1266, _--= 4017.