Note: Descriptions are shown in the official language in which they were submitted.
12412~34
NOVEL ANTEIRACYC:LTNE ANTIBIOTICS
BACKGROUND OF THE INVENTION
1. Field of the Invention
The present invention relates to novel anthracycline
antibiotics which are produced by microorganisms belonging
to the genus Streptomyces.
2. Description of the Prior Art
As anthracycline type antibiotics, daunomycin of
US. Patent 3,616,242) and adriamycin (cf. US. Patent
3,590,028~, obtained from a culture solution of actinomycetes
have been hitherto known. These compounds exhibit a broad
anti-cancer spectrum against experimental tumors and have
been widely utilized as chemotherapic agents against cancers
also from a clinical aspect. However, the anti-cancer
action of daunomycin and adri~mycin is not necessarily
satisfactory though they show a considerably potent action.
Thus, attempts to produce various compounds analogous
thereto have been made by various means of fermentation,
semi-synthesis, conversion using microorganisms and, some
anthracycline antibiotics have already been proposed (for
example, Published Examined Japanese Patent Application
3~915/76 aclacinomycins A and B), T. Ok et at, The Journal
of Antibiotics, vol. 33, pages 1331-1340, F. Armenia,
Topics in Antibiotic Chemistry, vol. 2, pages 102-279,
I ~Z~34
published by Ellis Howard Limited, Published Unexamined
Japanese Patent Application 56494/82 ~4-demethoxy-11-
deoxydaunomycin, etc.), Published Unexamined Japanese
Patent Application 15299~81 (rhodomycin series antibiotics),
etc. are disclosed).
As anthracycline antibiotics as anti-cancer agents,
a variety of analogous compounds have been proposed as
described above; a part of them has teen widely used for
clinical purpose and provided for clinical test.
However, none of them is satisfactory both in
toxicity and anti-cancer action. Further, results of anti-
tumor agents obtained with in vitro tests and animal tests
are not always reflective directly on anti-cancer action
against human and therefore, investigations are required
from various viewpoints. For this reason, it has been
desired Jo propose compounds belonging to a further new
class, with respect to anthracycline antibiotics which
have been evaluated in a way.
SWALLOWER OF THE INVENTION
According to the present invention there is provided a process
for preparation of a novel anthracycline antibiotic represented by
formula: O OH
YO-YO
HO o OH O
Ho 0
H NO
lZ~1284
wherein Y represents a group shown by:
OH
'
OH
10 SHEA H
OH or OH
in which a strain of the genus Streptomyces capable of a
producing said anthracycline compounds is cultivated in a
nutrient medium whereby said compounds are recovered from
the broth.
The present invention also provides a novel
anthracycline antibiotic obtained by the above process and
represented by the same general formula.
These compounds are antibiotics structurally
characteristic of ring A of the anthracyclinone skeleton.
Hereafter, among compounds shown by formula (I),
various antibiotics are listed. For instance, the
antibiotic represented by formula:
,~;,
. . I
~2412~4
O Ox COUCH
$
OH o OH O (I-a)
owe
OH NH2
is designated D788-6.
The preferred compounds according to the invention
are an antibiotic represented by formula:
- ~Z4~84
o OH OH
(I-b)
OH O OH O
H3C O
,
OH NH2
is designated D788-7;
an antibiotic represented by formula:
O OH ox
c)
C
.
HO NH2
is designated D788-8;
an antibiotic represented by formula:
124128~
OXXH3
O OH O
(I-d)
HO O OH
Ho
NH2
I
. .
is designated D788-9; and;
an antibiotic represented by formula:
O I COO OH
HO O OH O (I-e)
H3C1--g
HO NH2
- is designated D788-10.
These compounds exhibit a highly inhibitory action
against proliferation of cultured leukemia cell L 1210 and
are per so useful as anti-cancer agents.
1241284
Inhibitory action against proliferation of mouse L 1210
leukemia cell and against synthesis of nucleic act
L 1210 cells of S x 104/ml were inoculated on,
e.g., RPM 11640 medium (Rosewellberg Research Laboratories)
containing 20% bovine serum and, the substances of the
present invention were added thereto, respectively, in
a concentration of 0.02 to 0.25 gel in a similar fashion.
Cultivation was performed at 37C in a carbon dioxide
culture bottle. Then, a 50% inhibitory action against
proliferation was determined based on control fraction.
Further, the aforesaid L 1210 culture cell was suspended
in 10% bovine serum-containing RPM 11640 medium in a
concentration of S x 10 my After cultivating at 37~C
for 1 to 2 hours in a carbon dioxide culture bottle, the
substance was added in various concentrations and, 14C-
uridine (0.05 Somali or 14C-thymidine ~0.05 Somali) was
further added thereto 15 minutes after followed by culturing
at 37C for 60 minutes. A cold 10% trichloro-acetic acid
was added to the reaction solution to discontinue the
reaction and at the same time, matters insoluble in the
acid were precipitated. After washing further twice with
cold 5% trichloroacetic acid, the insoluble matters were
dissolved in formic acid and, radioactivity was measured.
A 50% intake inhibitory concentration was determined from
an intake rate of radioactivity, based on the control
-- 7 --
1241Z84
fraction to which no substance was added. The results
are shown in Table l.
Table l. Inhibitory Action of Compounds of
this Invention against Proliferation
of Mouse Leukemia L 1210 Culture Cell
and against Synthesis of Nucleic Acid
50% Inhibitory Concentration (ISSUE)
gel
,_ _
Inhibition Inhibition Inhibition
of Cell of DNA of RNA
Proliferation Synthesis Synthesis
D 788-6 0.25 2.6 1.30
D 788-7 0.0003 0.29 0.68
D aye 0.22 3~4 1.58
D 788-9 0.22 4.2 2.59
D Lowe 0.083 lo lo
The anti-tumor activity against mouse L 121Q
leukemia is shown in Tale 2.
124128~
Table 2. In viva Anti-Tumor Activity of D788-7
a ain't louse L 1210 Leukemia
g
Anti-Tumor Activity in viva
Dose
~g/k~/day)Anti-tumor Effect TIC
250 100
125 172
62.5 204
32 187
16 133
8 104
4 102
mice: CDFl n = 6
Inoculation: 1 10 cell/mouse (imp.)
Treatment: day 1 to day lo (imp.)
The foregoing ant~racycline antibiotics can be
produced by culturing in a medium comprising suitable
nutrient sources a strain producing the product of the
present invention which is easily isolated by a conventional
variation treatment of a strain belonging to the genus
Actinomyces and capable of producing daunomycin and
analogous compounds thereto which is isolated from the
soil or is known, using as a variant, for example, N-methyl-
124:~2~34
N'-nitro-N-nitrosoquanidine (NAG). Among these producing
strains, a specific example includes an RPM-5 strain which
is a variant obtained by a variation treatment of Streptomyces
D 788 strain producing daunomycin and paumycin, which is
newly isolated from the soil, with NAG.
This strain has been deposited on July 2, 1984 at
the Fermentation Research Institute, Agency of Industrial
Sciences and Technology under the international deposit
accession No. 811 (FEZ BP-811).
Hereafter, the bacteriological properties of the
RPM-5 strain are described below.
(i) Morphology
Linear aerial Muslim is developed from branching
substrate Mazola but no whirl is formed. Matured spores
in a chain of 10 or more have a diameter of about 0.6 to
0.8 x 0.9 to 2.5 microns. The spores have smooth surfaces.
Neither ascospore nor flagella spore is formed.
(ii2 Growth conditions in various media
With respect to color indication, standard shown
within parenthesis is based on "System of Color Wheels
for Streptomycete Taxonomy" written by Tresner & EDGY. Backup
(J. Apply Microbial., vol. 11, pages 335-338, 1963), which
is supplemented by "Color Standard" published by Nippon
Color Research Laboratories.
-- 10 --
~Z4~Z13~
(iii) Physiological characteristics
(1) Growth temperature range: (tested at each
temperature of 20C, 28C, 30C, 37C
and 42CC at pi 6.0 using yeast-maltose-
ajar medium) Grow at each temperature
of 20 to 37C but cannot grow at 42C.
I Liquefaction of gelatin:
positive (cultured at 20C using
glucosepeptone-gelatin medium)
to Hydrolysis of starch:
positive (starch-inorganic salt-agar medium)
(4) Coagulation of skim milk and peptonization:
all negative at the initial stage but 15
days after cultivation, peptonization
started.
(5) Formation of melanine-like pigment:
(using tryptone-yeast-broth, peptone-yeast-
iron-agar and tyrosine-agar media). positive
in all media
(iv) Utilization of various carbon sources: (Pridham-
Gottlieb ajar medium
- 1. L-Arabinose positive
2. D-Xylose positive
3. D-Glucose positive
4. D-Fructose positive
~241~8~
S. Sucrose positive
6. Instill positive
7. L-Rhamnose negative
8. Ruffians negative
9. D-Mannitol positive
Cultivation of producing bacteria relating to
the present invention can be performed in medium compositions
which are ordinarily used as nutrient sources for actinomycetes
and are per so known. For example, as carbon sources,
there can be used glucose, glycerine, sucrose, starch,
maltose, animal and vegetable oils, etc.; as nitrogen
sources, there can be used, for example, organic such as
soybean powders, meat extract, yeast extract, petunia,
corn steep liquor, cotton seed lees, fish powders, etc.
and inorganic nitrogens such as ammonium sulfate, ammonium
chloride, sodium nitrate, ammonium phosphate, etc.
If necessary and desirer sodium chloride, potassium
chloride, phosphates or bivalent metals salts such as My++,
Cay , Zen , Fe , Cut , on , or No , etc. and amino acids
or vitamins can be added. In addition, for purpose of
preventing foaming during fermentation, deforming agents,
for example, silicone (made by Shunts Kagaku CRY.,
-KM I trademark), etc. can be appropriately added.
Conditions for fermentation such as temperature,
phi aerial agitation and a time period for the fermentation,
- 12 -
lZ41Z~
etc. may be chosen in such a manner that the bacteria used
accumulates the maximum amount of the compound.
For example, it is advantageous to perform the fermentation
at temperatures of 20 to 40C, preferably at 28C, at pi
of S to q, preferably 6 to 7, for a time period for the
fermentation of 1 to 10 days, preferably 6 days.
To isolate and collect the substances D788-6 to 10
from the culture solution, toe culture solution after
completion of the fermentation is subjected to centrifugal
separation or filtered in the presence of a suitable
filtering aid such as diatomaceous earth thereby to
separate into the bacteria and the supernatant or the
filtrate. The substances are extracted from the supernatant
with organic solvents such as chloroform, Tulane, ethyl acetate,
etc. at pi of 7 to 9.
From the bacteria, the substances are extracted,
if necessary and desired, using organic solvents such as
acetone, methanol, ethanol, buttonhole, etc. Each of the
extracts is concentrated to dryness to obtain red, crude
powders. The powders are treated with chromatography
using adsorption carriers, for example, synthetic adsorbing
resins or silica gel, or, treated with anionic ion exchange
resins and cat ionic ion exchange resins, singly or in
suitable combination, to harvest each of substances D788-6
to 10 in a pure form.
- 13 -
~L241284
Hereafter, the present invention will be described
in more detail, referring to the examples below.
Example 1
- From an YE (0.3% yeast extract, 1% soluble starch,
1.5% ajar, pi 7.2) slant culture solution of Streptomyces
D 788, RPM-5 strain (deposited in Fermentation Research
Institute, Agency of Industrial Science and Technology
under the accession No 77031., a platinum loop was taken
out and inoculated in a 500 ml volume Erlenmeyer's flask
in which 100 ml Alcott of a seed culture medium described
below was charged and sterilized Shake culture was
performed at 20C for 2 days in a rotary shaker (220 rum)
to prepare a seed culture.
Seed culture medium:
Soluble starch 0.5 %
Glucose 0.5 %
Susan Meat soybean powder,
made by Ajinomoto Limited 1.0 %
Yeast extract 0.1 %
Sodium chloride 0.1 %
Potassium secondary phosphate 0.1 %
Magnesium sulfate (containing OWE) 0.1 %
Tap water
pi 7.4 (before sterilization
- 14 -
~2412~34
Then, 15 liters of a production medium having a
composition described below were charged in a 30 liter-
volume jar fermenter and then sterilized. Then, and,
750 ml (corresponding to 5%) each of the aforesaid seed
culture solution was added thereto and inoculated.
Production medium:
Taiwan yeast 5 %
Soluble starch 7.5 %
Yeast extract 0.2 g
Sodium chloride 0.2 %
Calcium carbonate 0.3 %
Mineral mixture* 0.06 %
Tap water
pi 8.2 (before sterilization)
A solution of 2.8 g of Quiz, 0.4 g of Phase
OWE, 3.2 g of Unyoke OWE and 0.8 g of ZnSO4 OWE
in 500 ml of distilled water.
When cultivation was performed at 28C for 130 hours
at an aerial amount of 15 loin while agitating at 45
rotations/min., the culture solution became a deep red
brown color because of the product. The culture solution
was collected from the jar fermenter. The pi of the culture
solution was rendered 1.7 with gone. sulfuric acid followed
- 15 -
~L241284
by agitation at room temperature for about 1 hour.
To the culture solution was added 2% of a filtering aid
and the bacteria was separated by filtration to obtain
13.5 liters of the filtrate. Further, the bacterial
fraction was suspended in 6 liters of acetone followed
by agitating for 20 minutes and extraction. After
filtering, the acetone extract was taken and concentrated
to about 1.5 livers under reduced pressure. The concentrate
was combined with the previously obtained filtrate to make
15 liters.
The aforesaid filtrate tpH was adjusted to 2.0 to
2.5 with ON sodium hydroxide was passed through a column
packed with 750 ml of Die Ion HP-20*~synthetic absorbing
resin, made by Mitsubishi Chemical Co., Ltd.2 at SO of 4.5
to absorb the product. Thereafter, washing with lo liters
of water having pi 1.7 toil. sulfuric acid was performed
and then the absorbed matter was eluded with 1.4 liters
of 50% acetone water (pi 1.7~. The equate was concentrated
under reduced pressure to about 800 ml. Then, the concern-
irate was adjusted to pi of 8.5 with ON sodium hydroxide
and extracted with chloroform (2 liters in total).
The thus obtained chloroform extract was washed with water
and then a saturated sodium chloride aqueous solution and
dried over Glabrous salt. After Glabrous salt was
filtered off, the filtrate was concentrated to a small
- trade mark
~-~ - 16 -
Jo .
12~1284
quantity under reduced pressure. An excess of Nixon
was added to cause precipitation. The filtrate was
collected by filtration and dried in vacuum to obtain
1.84 g of crude powders containing D7~8-6, D788-7, D788-8
and D~B8-9.
On the other hand, the residue remained after the
extraction with chloroform was adjusted to pi of 2.5 with
ON hydrochloric acid and then extracted with n-butanol
(1.5 liters in total. The thus obtained buttonhole extract
was rinsed with water showing pi of 2.5. After ~utanol
was removed by concentration under reduced pressure, the
residue was dissolved in a small quantity of methanol.
An excess of Tulane was added to the solution to cause
precipitation.- The solvent was removed by distillation
to obtain 5.0 of crude powders containing D788-10.
Example 2
n a column, 3Q g of awoke Silica Gel C-20Q jade by
Wake Junk Kiwi Cage Lid was packed with a chloroform-
methanol C20:1~ mixture. The crude powders, 600 my,
obtained by extraction with chloroform in Example 1 were
dissolved in a small quantity of a chloroform-methanol
I mixture and the solution was adsorbed to the upper
layer of the silica gel column followed by development
with the same solvent system. After aglycon eluded out
earlier was removed, development was performed with 160 ml
trade mark
- 17 -
12412~
of a chloroform-methanol (15:1) mixture, 140 ml of the
same (13:1) mixture and then 260 ml of the same (12:1)
mixture to elude a D788-6 fraction. Further, 200 ml of
a chloroform methanol (10:1) mixture was flown down to
elude a fraction containing D788-9. Then, elusion was
performed with 200 ml of a chloroform-methanol-water
(200 : 20 : 0.5~ mixture and then 200 ml of the same
~180 : 20 : 12 mixture to obtain a fraction containing
D~88-~ and D788-8. The weight of the crude powders in
each fraction obtained by concentration to dryness was
as follows.
D~88-6 Fraction 21 my
D788-7 Fraction 45 my
D~88-8 Fraction 79 my
D~88 9 Fraction 85 my
Each of the crude powders described above was
purified by thin layer chromatography for fractionation,
as described below.
D~88-6: The aforesaid D788-6 fraction was purified
using a silica gel thin layer ~2Q x 20 cam (PI 254 Silica
Gel, made by Mercy Ionic for fractionation. The fraction
was spotted in a horizontal line shape at a location of
15 mm from the lower end of the thin layer followed by
development with a chloroform-methanol-water (25 : 10 : 1)
trade mark
- 18 -
,,~ ,...
' .`-~-~ '
41Z8~
mixture. D788-6 bands were collected and extracted with
a chloroform-methanol (8:1) mixture. After the extract
was concentrated to dryness, the obtained powders were
dissolved in a Old acetic acid buffer solution (pi 3.0)
and the solution was extracted and washed with chloroform.
After the aqueous phase remained after the extraction was
adjusted with ON sodium hydroxide to pi of 8.5, the aqueous
phase was extracted with chloroform. After the resulting
extract was washed with water and then with a saturated
sodium chloride aqueous solution, it was dried over
Glabrous salt. The chloroform extract was filtered and
the filtrate was concentrated under reduced pressure.
An excess of Nixon was added to the concentrate to
cause precipitation. The product was collected by
filtration and dried in vacuum to obtain 10 my of pure
D788-6 substance.
D788-7: The aforesaid D788-7 fraction was subjected
to thin layer chromatography of silica gel Shapiro) for
fractionation using development solvent, chloroform-methanol-
water-acetic acid ~120 : 25 : 6 : 141. Bands corresponding
to D~88-J were collected and extracted with a chloroform-
methanol-water (4 : 1 : 0.51 mixture. After the extract
was concentrated to dryness, the residue was dissolved in
lo acetic acid buffer (pi 3.01 and the solution was
washed with chloroform and extracted with chloroform at
-- I --
1241Z84
pi of 8.5 in a manner similar to the purification of
D788-6 described above to give 21 kg of pure D788-7
substance.
D788-8: The aforesaid D788-8 fraction obtained
by silica gel column chromatography was purified by thin
layer chromatography for fractionation (swooper) using
development solvent, chloroform-methanol-water-acetic
acid-28% ammonia water (125 : 55 : 5.5 : I : 1.1).
Bands corresponding to D788-8 were collected and extracted
with a chloroform-methanol-water (4 : 1 : 0.5) mixture.
Subsequently, the extract was treated in a manner similar
to the purification of the D788-~ fraction described
above to it 28 my of pure D788-8.
D788-9: The D~88-~ fraction (8Q my) obtained by
silica gel chromatography described above was subjected
to a column filled up with a suspension of 4 g of Wake
Silica Gel C-2Q0 (swooper in a chloroform-methanol (20:1)
mixture and again subjected to chromatography developing
with the same solvent system to obtain D788-9 having a
purity of about 50~. This was further subjected to thin
layer chromatography for fractionation (swooper) and developed
with development solvent, a chloroform-methanol-water-
acetic acid ~120 : 25 : 6 : 14~ mixture to separate and
purify. Bands corresponding to D788-9 were collected and
extracted with a cnloroform-methanol-water (4 : 1 : 0.5)
mixture. Subsequently, the extract was treated in a
_ 20 -
lZ4128~
manner similar to D788-6 descried above to give 22 my
of pure D788-9.
Example 3
The crude powders (5 g) extracted with buttonhole
containing D788-10 obtained in Example 1 were subjected
to a column prepared by filling up with a suspension of
35 g of awoke Silica Gel C-200 swooper in chloroform-
methanol-water (100 : 10 : 0.5) and then developed, in
swoons, with 200 ml of a chloroform-methanol-water
(90: 10: 1) mixture, 360 ml of the same (80 10: 1)
mixture, 240 ml of the same (70: 10: 1) mixture and
then 210 ml of the same (60: 10 1) mixture to obtain
further purified D788-10 fractions. The D788-10 fractions
were collected and concentrated under reduced pressure
to obtain partly purified D788-10 powders. The powders
were dissolved in a diluted sodium hydrogen carbonate
aqueous solution. After the solution was washed with
chloroform and the remained aqueous phase was adjusted
to pi of 2.5 with ON hydrochloric acid, the aqueous phase
was extracted with n-butanol. The extract was concentrated
under reduced pressure to obtain 80 g of powders.
The powders were further purified by thin layer chromatography
for fractionation using development solvent, chloroform-
methanol-water acetic acid-28% ammonia water (125: 55:
5.5 : 0.9 : 1.1). Fractions corresponding to D788-10 were
-- 21 --
124~ 84
collected and extracted with a mixture obtained by adding
a drop of acetic acid to chloroform-methanol-w~ter (4: 1
0.5). After the extract was concentrated under reduced
pressure, the residue was dissolved in a diluted sodium
hydrogen carbonate aqueous solution. After the solution
was washed with chloroform, the solution was adjusted to
pi of 2.5 with ON hydrochloric acid and then extracted
with n-butanol. The extract was concentrated to dryness
to obtain 32 my of purified D788-10 substance.
Physico-c~emical properties of each of the substances
obtained in the above examples are shown below.
D788-6
A) melting point 138 - 140C (decomposed
By ICES I 165 (c=Q.015, methanol
C) W and visible absorption spectra (methanol)
Max nut Mel% ): 206 (-3111
235 ~647)
255 (403)
290 ~141)
492 ~229
Shea ~177)
Shea (153~
Do IT absorption spectrum (KBr~ y cm 1
1720, 1590, 1450, 1420, 1390, 1280, 1~70,
1190, 1160, 1000, 980
-- 22 --
~L24:1Z84
E) Len spectrum (CDC13) Pam
1.1 (OH, t, J=7.5 H-14)
1.33 (OH, d, J=6.5 H-6')
1.51 (lo, dud, J=12.4 Howe)
1.6-2.1 (OH, m, H-13, Hub
2.1-2.4 (OH, m, H-8)
3.05 (lo, dud, J=11.4 H-3')
3.41 (lo, by, H-4')
3.69 (OH, s, COUCH )
4.1 (lo, q, J=6.5 H-5')
4.26 (lo, s, H-10)
5.22 (lo, by, H-7 3
5.43 (lo, by, H-l')
7.22 (lo, dud, J=8~2 H-3 )
7.62 (lo, t, J=8H-2 )
7.71 (lo, dud, J=8.2 Hal )
D788-7
A) melting point 165 - 167C
B) Do 16 (c=0.0125, methanol)
C) W and visible absorption spectra (in methanol)
SHEA no (E ) :207 (332)
Max lcm235 (702)
254 (424)
292 (139)
492 (254)
Shea (187)
528 (169)
- 23 -
~2~1284
D) IT absorption spectrum (KBr) yam
1595, 1450, 1430, 1400, 1290, 1230,
1190, 1160, 1110, 1005, 980
E) lH-NMR spectrum (CDC13 + CD30D) Pam
1.09 (OH, t , J=7.5 H-14)
1.32 (OH, d , J=6.5 H-6')
1.6-1.9 (OH, m , H-2', H-13)
2.2 (OH, by, H-8 )
2.7-3.2 (lo, m , H-3')
3.46 (lo, by, H-4')
4.12 (lo, q , J=6.5 H-5')
4.81 (lo, s , H-10)
5.1 (lo, by, H-7
5.4 (lo, by, H-l')
7.23 (lo, dud, Jo 1.5 H-3)
7.64 (lo, t Jo H-2 )
7.79 lo dud, Jo 1.5 H-l)
D788-8
A) melting point 126-128C (decomposed)
B) Do + 309 (c=0.0175, methanol)
C) US and visible absorption spectra (in methanol)
_ 24 -
~241Z~34
SHEA no (Elm) : 206 (owe)
269 (619)
494 (283)
515 (300)
Shea (183)
D) IT adsorption spectrum (RBr) y cm 1
1600, 1460, 1400, 1370, 1285, 1250,
1~00, 1170, 1110, lQlS, 980
F.) lH-NMR spectrum (CDC13-CD30D) Pam
1.3 (lo, d, J=6.5 H-6')
1.4 (OH, d, J=6.5 H-14)
1.5-1.9 OH m, Ho
2.5 (lo, dud, J=18, 4.5 Howe)
2.8 (lo, d, J=18 Hub
2.8-3.2 (lo, m, H-3')
3.38 (lo, by, H-4')
3.98 lo q, J=6.5 H-5')
4.49 (lo, q, J=6.5 H-13)
5.2 (lo, by, H-l')
6.97 (lo, by, H-10)
7.18 (lo, dud, Jo 1.5 H-3)
7.59 lo t, Jo H-2 )
7.75 (lo, dud, Jo 1.5 H-l)
- 25 -
lZ4~84
D788-9
A) melting point 175-1~0C
B) [~]25 + 62 (c=0.0145, methanol)
C) W and visible absorption spectra (in methanol)
kiwi Noel ) : 206 13677'
255 (428)
291 (153)
Shea (212)
493 (226)
Shea (171)
526 (1~8)
D) IT absorption spectrum ~KBr) y cm 1
1725, Shea, 1600, 1450, 1430, 1400,
1290, 1240, 1190, 1165, 1110, 1005, 980
E) H-NMR spectrum (CDC13) Pam
1.36 (OH, d , J=6.5 H-6')
1.5-2.0 (OH, m , H-2')
2.2 (lo, dud, J=16, 4.5 Howe)
2.27 (OH, s , H-14)
2.6 (lo, d , J=16 Hub
2.9-3.3 (lo, m , H-3')
3.46 (lo, by, H-4')
3.69 (OH, s , COUCH)
4.13 (lo, q , J=6.5 H-S')
4.37 (lo, s , H-10)
5.12 (lo, d , J=4.5 H-7)
~2~1289~
5.42 (lo, by, H-l')
7.23 (lo, dud, Jo lo H-3)
7.6 (lo, t , Jo H-2)
7.77 (lo, dud, Jo 1.5
D-788-10
A) melting point 174-176C
B) tall 25 + 200 (coo. 0185 r methanol)
C) W and visible absorption spectra yin methanol)
SUE m(Elcm) 2296 (3761)
257 (32g)
497 (169)
Shea (155)
D) IT absorption spectrum (KBr) cm
1710, 1600, 1450, 1390, 1280, 12~0,
1190, 1160, 1115, 101~, 980
E) ENAMOR spectrum (CDC:13-CD30D) Pam
1.3 (OH, d, J=6.5 H-6')
1.4 (OH, d, J=6.5 H-14)
1.5-1.9 (OH, m , H-2')
2.1-2.8 (OH, m , H-8
3.7 (lo, by, H-4')
4.0 (lo, q, J=6.5 H-13)
4.1 (lo, s , H--10)
-- 27 --
5.1 (lo, by, H-7 )
5.5 (lo, by, H-l )
7.2 (lo, dud, Jo 1.5 H-3)
7.6 (lo, t , Jo H-2 )
7.7 lo dud, Jo 1.5 H-l)
While the invention has been described in detail
and with reference to specific embodiments thereof, it
will be apparent to one skilled in the art that various
changes and modifications can be made therein without
departing from the spirit and scope thereof.
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