U.S. patent application number 16/191007 was filed with the patent office on 2019-05-30 for novel harringtonines salts in the crystalline state, their use for the purification of the corresponding drug substance and as c.
The applicant listed for this patent is Thierry BATAILLE, Julie BLANCHARD, Nina RADOSEVIC, Jean-Pierre ROBIN, Thierry ROISNEL. Invention is credited to Thierry BATAILLE, Julie BLANCHARD, Nina RADOSEVIC, Jean-Pierre ROBIN, Thierry ROISNEL.
Application Number | 20190161493 16/191007 |
Document ID | / |
Family ID | 52278654 |
Filed Date | 2019-05-30 |
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United States Patent
Application |
20190161493 |
Kind Code |
A1 |
ROBIN; Jean-Pierre ; et
al. |
May 30, 2019 |
NOVEL HARRINGTONINES SALTS IN THE CRYSTALLINE STATE, THEIR USE FOR
THE PURIFICATION OF THE CORRESPONDING DRUG SUBSTANCE AND AS
CHEMOTHERAPEUTIC AGENTS GIVEN ALONE OR COMBINED WITH RADIOTHERAPY
OR AS IMMUNOMODULATING AGENTS
Abstract
The present invention concerns harringtonines salts never
described in the crystalline state exhibiting a protonated nitrogen
seen in solid state analysis and having general formula 1,
##STR00001## comprising solvates, made by reacting a cephalotaxine
ester alkaloid base having formula 2, ##STR00002## in which R.sup.1
is, but not limited to, alkyl, aryl, cycloalkyl, heteroalkyl,
heteroaryl or heterocycloalkyl, and R.sup.2 is, independently, but
not limited to H, alkyl, aryl, cycloalkyl, heteroalkyl, heteroaryl
or heterocycloalkyl, with an acid having general formula AH in a
non-aqueous crystallization solvent, wherein said salt has a large
water solubility. The invention also relates to a process for
preparing and purifying these salts and their use as
chemostherapeutic drugs, alone or combined with radiotherapy, or as
immunomodulating agents.
Inventors: |
ROBIN; Jean-Pierre; (Geneve,
CH) ; RADOSEVIC; Nina; (Geneve, CH) ;
BLANCHARD; Julie; (Puyricard, FR) ; ROISNEL;
Thierry; (Thorigne-Fouillard, FR) ; BATAILLE;
Thierry; (Thorigne-Fouillard, FR) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
ROBIN; Jean-Pierre
RADOSEVIC; Nina
BLANCHARD; Julie
ROISNEL; Thierry
BATAILLE; Thierry |
Geneve
Geneve
Puyricard
Thorigne-Fouillard
Thorigne-Fouillard |
|
CH
CH
FR
FR
FR |
|
|
Family ID: |
52278654 |
Appl. No.: |
16/191007 |
Filed: |
November 14, 2018 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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15109254 |
Jun 30, 2016 |
10150776 |
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PCT/EP2014/079456 |
Dec 30, 2014 |
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16191007 |
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61941723 |
Feb 19, 2014 |
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61922248 |
Dec 31, 2013 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P 35/00 20180101;
C07D 491/147 20130101; C07D 491/14 20130101 |
International
Class: |
C07D 491/147 20060101
C07D491/147; C07D 491/14 20060101 C07D491/14 |
Claims
1. A harringtonines salt in the crystalline state exhibiting a
protonated nitrogen seen in solid state analysis and having formula
1, ##STR00022## comprising solvate, made by reacting a
cephalotaxine ester having formula 2, ##STR00023## in which R.sup.1
is, but not limited to, alkyl, aryl, cycloalkyl, heteroalkyl,
heteroaryl or heterocycloalkyl, and R.sup.2 is, independently, but
not limited to H, alkyl, aryl, cycloalkyl, heteroalkyl, heteroaryl
or heterocycloalkyl, with an acid having general formula AH in a
crystallization solvent, wherein the said salt has a water or
alkohol solubility ranged approximately from 5 mg/mL to
approximately 100 mg/Ml.
2. The salt of claim 1 wherein the cephalotaxine ester reactant is
homoharringtonine (=omacetaxine) having formula 2 in which R.sup.2
is hydrogen and R1 have below formula 3. ##STR00024##
3. The salts of claim 1 wherein the acid is an organic acid but not
limited to, selected among the following list: fumaric, maleic,
citramalic, malic, tartaric, tartronic, succinic, itaconic, citric
acid or salicylic acid.
4. The salts of claim 1, having below formula ##STR00025## in which
the malic acid is of configuration 2S having formula
##STR00026##
5. The salts of claim 4, wherein the malic acid is of configuration
2R having formula ##STR00027##
6. The salt of claim 1, named (3S,4S,5R,2'R)-homoharringtonine
hydrogen (R)-malate exhibiting the below formula: ##STR00028##
7. The salt of claim 1, named (3S,4S,5R,2'R)-homoharringtone
hydrogen succinate exhibiting the below formula: ##STR00029##
8. The salt of claim 1, named (3S,4S,5R,2'R)-homoharringtonine
hydrogen (2'''S,3'''S)-tartrate exhibiting the below formula:
##STR00030##
9. The salt of claim 1, named (3S,4S,5R,2'R)-homoharringtonine
hydrogen (2'''R,3'''R)-tartrate exhibiting the below formula:
##STR00031##
10. The salt of claim 1, named (3S,4S,5R,2'R)-homoharringtonine
hydrogen itaconate exhibiting the below formula: ##STR00032##
11. The salt of claim 1, named (3S,4S,5R,2'R)-homoharringtonine
hydrogen fumarate exhibiting the below formula: ##STR00033##
12. The salt of claim 1, named (3S,4S,5R,2'R)-homoharringtonine
hydrogen tartronate exhibiting the below formula: ##STR00034##
13. The salt of claim 1, named (3S,4S,5R,2'R)-homoharringtonine
hydrogen malonate exhibiting the below formula: ##STR00035##
14. The salt of claim 1, named (3S,4S,5R,2'R)-homoharringtonine
dihydrogen citrate exhibiting the below formula: ##STR00036##
15. The salt of claim 1, named (3S,4S,5R,2'R)-homoharringtonine
salicate exhibiting the below formula: ##STR00037##
16. The salt of claim 1 as crystalline form comprising solvate and
co-crystal.
17. The cation (3S,4S,5R,2'R)-homoharringtoninium as described in
FIGS. 2.3.1, 2.4.1, 2.5.1, 2.6.1, 2.8.1, 2.9.1, 2.11.1, and 2.12.1,
exhibiting the below formula ##STR00038##
18. The process of preparation and purification of salts of claim 1
comprising contacting a natural, hemi-synthetic or synthetic
harringtonine or its semi-synthetic analog with a weak acid in
suspension or in solution in a suitable non-aqueous solvent,
preferably an alcohol or mixed at the solid state either at the
amorphous state or at the crystalline state then recrystallized
said salt in a suitable non aqueous solvent, preferably an alcohol,
the said process being also when repeated a method of purification
including enantiomeric (fractional crystallization).
19. The process of claim 18 wherein the harringtonine is
homoharringtonine having formula represented in claim 2.
20. The process of claim 18 wherein the harringtonine is an
harringtonine analog having general formula: ##STR00039## in which
R.sup.1 is, but not limited to, alkyl, aryl, cycloalkyl,
heteroalkyl, heteroaryl or heterocycloalkyl, and R.sup.2 is,
independently, but not limited to H, alkyl, aryl, cycloalkyl,
heteroalkyl, heteroaryl or heterocycloalkyl.
21. The process of claim 18, wherein the acid is, but not limited
to, selected among the following list: fumaric, maleic, citramalic,
malic, tartaric, tartronic, succinic, itaconic, salicylic or citric
acid.
22. A pharmaceutical dosage form comprising a pharmaceutically
acceptable carrier and a therapeutically effective amount of a
mineral or organic salt or solvate or co-crystal of claim 1.
23. A method of treatment comprising administering a
therapeutically effective amount of a pharmaceutical dosage form of
claim 22 to a patient or an animal suffering from cancer including
their metastasis, leukemia, lymphoma, parasitic disease, ocular
proliferation and/or immune disorder and/or from viral disease.
24. A method of treating cancer, leukemia and/or lymphoma,
comprising administering to a patient or an animal in need thereof
the pharmaceutical dosage of claim 22, said pharmaceutical dosage
being administered alone or in combination with at least another
chemotherapeutic agents, eventually combined with radiotherapy.
25. The method of claim 24, wherein the leukemia is acute myelod
leukemia (AML), myelodysplastic syndrome (MDS) and
myeloproliferative disorders including chronic myelogenous
leukemia, polycythemia vera, essential thrombocythemia,
myelosclerosis, and wherein the lymphoma is a multiple myeloma, a
Hodgkin disease or a Burkitt lymphoma, and wherein the cancer is a
breast cancer, a brain cancer or a lung cancer.
26. The method of claim 25, wherein the breast cancer is a triple
negative breast cancer (TNBC).
27. The method of claim 25 wherein the brain cancer is a
neuroblastoma.
28. The method of claim 25, wherein the lung cancer is a non small
cell lung cancer (NSCLC).
29. A method for treating autoimmune disorder, comprising
administering to a patient or an animal in need thereof the
pharmaceutical dosage of claim 22, said pharmaceutical dosage being
administered alone or in combination with at least another
chemotherapeutic agent.
30. The method of claim 29, wherein the autoimmune disorder is a
systemic lupus erythematosus (SLE), a dermatomyositis, a psoriasis
or a lichen planopilaris (LPP).
31. The method of claim 30, wherein the lichen planopilaris (LPP)
is a frontal fibrosis alopecia (FFA).
Description
[0001] The present invention concerns crystalline salts of
harringtonines, protonated on their alkaloid nitrogen, definite by
their solid state analysis patterns, their process of preparation
allowing their use as drug substance for blending alone or in
combination in pharmaceutical composition useful as
chemotherapeutic agents, given alone or combined with radiotherapy,
or useful for treating parasitic and viral diseases and/or as
immunomodulating agents, particularly in using oral or parenteral
modes of administration.
[0002] Among harringtonines, homoharringtonine (=HHT, named
omacetaxine D.C.I. as drug substance) is a natural ester of
cephalotaxine (see scheme 1 and table 1), an alkaloid of
Cephalotaxus harringtonia, a rare and endangered Asian conifer
belonging to the Cephalotaxaceae family. HHT content in renewable
parts of Cephalotaxus is about a few dozen of mg only per kilo of
dry plant material. This characteristic, in despite of considerable
efforts performed by the U.S. National Cancer Institute, hampered
clinical development of omacetaxine for more than thirty-years. On
March 1998, the discovering of a new hemi-synthetic process by the
Applicant, allowed industrial production of homoharringtonine at
the kilo scale (U.S. Pat. No. 6,831,180 and Robin et al. Tet. Lett.
1999. p.2931)] and divided by 70 the need of rare plant material
(Nicolini et al., Leukemia Research, 2014, 38, p.11545).
[0003] Important Note:
[0004] It should be noted that chemical structure of hemi-synthetic
omacetaxine is strictly identical to the natural one version:
omacetaxine is not a semi-synthetic derivative as indicated in some
article published in literature (see scheme 1 and table 1). All
denominations of omacetaxine (OMA) or homoharringtonine (HHT)
included in this document are strictly equivalent regarding
molecular structure. The sentence "omacetaxine is a semi-synthetic
derivative of cephalotaxine" encountered in literature, is totally
devoid of scientific significance: the semi-synthetic appellation
suggests that a moiety of the molecule (cephalotaxine) would
natural and that the other moiety (the side chain) would be
unnatural (man designed) while the latter is strictly natural. When
only a portion of a molecule was produced by synthesis, the process
name is hemi-synthesis and the molecule is sometimes also called
hemi-synthetic.
[0005] Short History of Recent Development of
Homoharringtonine.
[0006] Initially, all above esters of cephalotaxine were discovered
by U.S. teams (Powel et al., J. Pharm. Sc., 1972, 61, p.1227) and a
large development program was performed by the United States
National Cancer Institute (Suffness et al., J. Nat. Cancer Inst.,
1988, 80, p.1095). In October 2012, the United States Food and Drug
Administration (FDA) granted accelerated approval for omacetaxine
mepesuccinate for the treatment of adult patients with chronic or
accelerated phase chronic myeloid leukemia (CML) who failed to
respond to two or more tyrosine kinase inhibitors (TKIs) [ref fda].
Since this approval, hundreds of articles or reviews related to
OMA/HHT were published in literature (more than 400 articles listed
in SciFinder database). Definitive approval of OMA was granted in
2014 (Alvandi et al., The Oncologist, 2014, 19, p 94). This
occurred after a very long and tumultuous period of clinical
development (Kantarjian et al., Clin. Lymph. Myel. Leuk. 2013 p.
530), including early clinical development of HHT and, to a lesser
extent, its congeners harringtonine (HA) and deoxyharringtonine
(DHA) in various institution in the U.S. and in China. Finally, the
successive involvement of seven pharmaceutical companies
(Vivorex/American Bioscience; Oncopharm; Stragen; Chemgenex;
Cephalon; TEVA) dispatched in 5 countries occurred before approval
of omacetaxine! More than 50 clinical trials in USA, China and
France involving more than 2000 patients.
[0007] Definition (See Scheme 1 and Table 1)
[0008] Homohamingtonine/Omacetaxine
Mepesuccinate/Synribo/Myelostat
[0009] The INN (Intemational Non-proprietary Name) "omacetaxine
mepesuccinate" (OMA) is a name reserved for homoharringtonine HHT
drug substance dedicated for pharmaceutical and medicinal use
regardless its natural, hemi-synthetic or synthetic origin [formely
named homoharringtonine]. Synribo (TEVA) and Myelostat (Oncopharm
corporation) are trademark (F-D-C Reports, Pharmaceutical Approvals
Monthly, 2001, 6, p.35).
[0010] Cephalotaxanes Including Numbering
[0011] Cephalotaxanes are particular alkaloids to date exclusively
extracted from the Cephalotaxaceae family which exhibit the
structural formula 1. Several substituants may be encountered on
this core structure: hydroxyl, ether, acyloxy etc. The eventual
presence of some additional double bound or intramolecular bridge
achieve to definite cephalotaxanes. Cephalotaxines 2 are
cephalotaxanes without acyloxy side-chain.
[0012] Cephalotaxine 2a and drupacine 2b are example of
cephalotaxines. Harringtonines 5 are particular cephalotaxanes
formed by attachment of a branched .alpha.-hydroxyacyloxy
side-chain at the 3-position of various cephalotaxines moieties.
Cephalotaxines 2 and harringtonines 5, are examples of
cephalotaxanes. Several dozen of cephalotaxanes have been isolated
from various Cephalotaxus species. 4 is the generic formula of
cephalotaxine esters (Takano et al., Phytochemistry, 1997, 44, p.
735 and cited references).
[0013] Harringtonines 5 (i. e. harringtonine=HA and
homoharringtonine=HHT) are particular cephalotaxine esters.
Cephalotaxine and its natural ester are gathered under the generic
term of cephalotaxane.
[0014] Harringtoids are semi-synthetic derivatives of
harrintonines.
[0015] Harringtonic acids are side-chain of harringtonines.
TABLE-US-00001 TABLE 1 NATURAL AND SEMI-SYNTHETIC ESTERS OF
CEPHALOTAXINE # Trivial name R.sub.2 R.sub.3 R.sub.4 Note #
Activity* 5a harringtonine (CH.sub.3).sub.2COH--(CH.sub.2).sub.2--
Me H (1) anticancer 5b homoharringtonine
(CH.sub.3).sub.2COH--(CH.sub.2).sub.3-- Me H (2) anticancer 5c
norharringtonine (CH.sub.3).sub.2COH--CH.sub.2-- Me H (3) none 5d
deoxyharringtonine (CH.sub.3).sub.2CH--(CH.sub.2).sub.2-- Me H (4)
anticancer 5e bishomoharringtonine
(CH.sub.3).sub.2COH--(CH.sub.2).sub.4-- Me H (5) none 5f
isoharringtonine (CH.sub.3).sub.2CH(CH.sub.2).sub.2-- Me OH (6)
none 5g neoharringtonine C.sub.6H.sub.5--CH.sub.2-- Me H (7)
cytotoxic 5h harringtonines R.sub.2-- Me R.sub.4 (8) N/A 5i
harringtoids R.sub.2-- R.sub.3 R.sub.4 (9) cytotoxic
[0016] (1) The first cephatotaxine ester isolated from oephalotaxus
harringtonia *In cancer area, for definition of term see [Suffness
et al in Journal of Natural Products 1982 p 1 Current Status of the
NCI Plant and Animal Product Program] CYTOTOXICITY is toxicity to
tumor cells in culture; ANTITUMOR is in vivo activity in
experimental systems; ANTINEOPLASTIC or ANTICANCER are the reserved
terms for reported clinical trials data.
[0017] (2) "Homo" means one more carbon than harringtonine; Named
omacetaxine (D.C.I.) as active pharmaceutical ingredient
[0018] (3) "nor" means one more less carbon.
##STR00003## ##STR00004##
[0019] Two haringtonines are very promising drugs in the treatment
of certain leukemia such as Chronic Myelogenous Leukemia (CML).
Both homoharringtonine and harringtonine were used in human
chemotherapy of leukemia for 30 years (see above Suffness et al.)
and a large number of semi-synthetic analogs such as 5 on scheme 1
were synthesized (see "5.2 Cephalotaxus Esters With Side Chain
Analogs" in above cited reference of Dumas et al.).
[0020] Surprisingly, never crystalline salts of harringtonines have
been isolated and described in literature.
[0021] However, in spite of the progress recorded in production,
purification and therapeutic use of homoharringtonine, several
disadvantages persist:
[0022] i) The cost of treatment for omacetaxine (Synribo) is
prohibitive: $28,000 for induction, $14,000 for monthly
treatments), this give about 180.000 $ per year, per patient
[Kantarjian et al. Journal of Clinical Oncology, 2013, p3600; Hagop
Kantarjian, personal communication]
[0023] ii) The use of the parenteral route of administration even
retards the development of this drug
[0024] iii) Preparation of formulations for parenteral use is
complicated by the use of lyophilization
[0025] iv) Formation of non-crystalline salts of harrintonines give
not as accurately defined compound as crystalline salts
[0026] v) There is some local intolerance to this product when
administered subcutaneously
[0027] vi) On the other hand, although it has been known for almost
40 years, there is still a slight doubt regarding the absolute
configuration of this series of natural product.
[0028] Recent Scientific Discovering Regarding Mechanism of
Activity of Harringtonines
[0029] The team of Steitz (Journal of Molecular Biology 2009, 389,
p. 146) recently demonstrated that homoharringtonine when in place
in its active site was protonated in a neutral media, implying that
alkaloid nitrogen protonation is imperative condition for the
manifestation of the activity of this ligand.
[0030] In addition, the team of Takano et al [J. Org. Chem. 1997 p.
8251) demonstrated experimentally that when the nitrogen lone pair
of homoharringtonine was occupied by an oxygen atom, the cytotoxic
activity was divided by a factor of at least 50. The authors
conclude that "the nitrogen lone pair on the cephalotaxine skeleton
appears to be essential for its activity".
[0031] The above mentioned team of Steitz showed that the absolute
configuration of homoharringtonine deposited in the Cambridge
Structural Database seems to be the opposite of that commonly
adopted in the literature.
[0032] The present invention relates to overcome the problems
mentioned above. It also demonstrated that the absolute
configuration in the deposited homoharringtonine Cambridge
Structural Database seems to be the opposite of that commonly
retained in the literature.
[0033] The eight example of single crystal X-ray diffraction of
homoharringtonine salt exhibited in FIGS. 2.3.1, 2.4.1, 2.5.1,
2.6.1, 2.8.1, 2.9.1, 2.11.1 and 2.12.1 clearly indicates that the
alkaloid moiety was efficiently protonated by the processes
described in the present invention. Moreover, the shortest distance
between said proton carried by the nitrogen is close to two
angstroms, showing the reality of the formation of a salt and not a
mere co-crystal.
##STR00005##
[0034] The present invention relates to overcome the problems
mentioned above, namely: [0035] raise doubt on the absolute
configuration of harringtonines [0036] provide a method of
administration of harringtonines protonated on their nitrogen
atom
[0037] As detailed above, the fact that the real active form of
harringtonines would be their nitrogen-protonated version was
recently supported by the above cited works of Seitz et al. and
Takano et al.
[0038] The present invention concerns novel water soluble
crystalline salts of homoharringtonine and their use as new
chemical entities for the formulation of new cancer
chemotherapeutic, or immunomodulating or antiparasitic agents and
to implement new processes for purification including enantiomeric
and determine the absolute configuration of the series.
[0039] The present invention describes the preparation of
crystalline salts of harringtonines as nitrogen-protonated form,
stable and soluble in water and their use for the manufacture of
pharmaceutical composition useful in the treatment of cancers,
leukemias, immune disease and as reversal agents.
[0040] The present invention describes an unambiguously proved
method of protonation of harringtonine nitrogen.
[0041] The present invention provides salts of harringtonines in
the crystalline state, protonated on their alkaloid nitrogen,
definite by their solid state analysis patterns, their process of
preparation from harringtonines and commercial organic acid
allowing their use as drug substance for blending alone or in
combination with other chemotherapeutic agents such as, but not
limited to, cytarabine or interferon or imatinib mesylate or
dasatinib or arsenic trioxide or all-trans-retinoic acid, in a
pharmaceutical composition particularly useful for treatment of
cancer, alone or combined with radiotherapy, in using oral or
parental modes of administration.
BRIEF DESCRIPTION OF DRAWINGS
[0042] FIG. 1.1 is infra-red (IR) spectrum of Homoharringtonine
(base alkaloid). FIG. 1.1 (i) is IR(ATR) spectrum in the
crystalline state. FIG. 1.1 (ii) is IR(ATR) spectrum in the
amorphous state.
[0043] FIG. 1.2 is infra-red (IR) spectrum of Homoharringtonine
hydrogen (S)-malate in the solid state. FIG. 1.2 (i) is IR(ATR)
spectrum in the crystalline state. FIG. 1.2 (ii) is IR(ATR)
spectrum in the amorphous state (film).
[0044] FIG. 1.3 is infra-red (IR) spectrum of Homoharringtonine
hydrogen (R)-malate in the solid state. FIG. 1.3 (i) is IR(ATR)
spectrum in the crystalline state. FIG. 1.3 (ii) is IR(ATR)
spectrum in the amorphous state (film).
[0045] FIG. 1.4 is infra-red (IR) spectrum of Homoharringtonine
hydrogen (2S, 3S)-tartrate in the solid state. FIG. 1.4 (i) is
IR(ATR) spectrum in the crystalline state. FIG. 1.4 (ii) is IR(ATR)
spectrum in the amorphous state (film).
[0046] FIG. 1.5 is infra-red (IR) spectrum of Homoharringtonine
hydrogen (2R, 3R)-tartrate in the solid state. FIG. 1.5 (i) is
IR(ATR) spectrum in the crystalline state. FIG. 1.5 (ii) is IR(ATR)
spectrum in the amorphous state (film).
[0047] FIG. 1.6 is infra-red (IR) spectrum of Homoharringtonine
hydrogen (2'''S)-citramalate in the solid state. FIG. 1.6 (i) is
IR(ATR) spectrum in the crystalline state. FIG. 1.6 (ii) is IR(ATR)
spectrum in the amorphous state (film).
[0048] FIG. 1.7 is infra-red (IR) spectrum of Homoharringtonine
hydrogen (2'''R)-citramalate in the solid state. FIG. 1.7 (i) is
IR(ATR) spectrum in the crystalline state. FIG. 1.7 (ii) is IR(ATR)
spectrum in the amorphous state (film).
[0049] FIG. 1.8 is infra-red (IR) spectrum of Homoharringtonine
succinate. FIG. 1.8 (i) is IR(ATR) spectrum in the crystalline
state. FIG. 1.8 (ii) is IR(ATR) spectrum in the amorphous state
(film).
[0050] FIG. 1.9 is infra-red (IR) spectrum of Homoharringtonine
hydrogen itaconate in the solid state. FIG. 1.9 (i) is IR(ATR)
spectrum in the crystalline state. FIG. 1.9 (ii) is IR(ATR)
spectrum in the amorphous state (film).
[0051] FIG. 1.10 is infra-red (IR) spectrum of salt named
homoharringtonine hydrogen fumarate in the solid state. FIG. 1.10
(i) is IR(ATR) spectrum in the crystalline state. FIG. 1.10 (ii) is
IR(ATR) spectrum in the amorphous state (film).
[0052] FIG. 1.11 is infra-red (IR) spectrum of Homoharringtonine
hydrogen tartronate in the solid state. FIG. 1.11 (i) is IR(ATR)
spectrum in the crystalline state. FIG. 1.11 (ii) is IR(ATR)
spectrum in the amorphous state (film).
[0053] FIG. 1.12 is infra-red (IR) spectrum of Homoharringtonine
malonate in the solid state. FIG. 1.12 (i) is IR(ATR) spectrum in
the crystalline state. FIG. 1.12 (ii) is IR(ATR) spectrum in the
amorphous state (film).
[0054] FIG. 1.13 is infra-red (IR) spectrum of Homoharringtonine
dihydrogen citrate in the solid state. FIG. 1.13 (i) is IR(ATR)
spectrum in the crystalline state. FIG. 1.13 (ii) is IR(ATR)
spectrum in the amorphous state (film).
[0055] FIG. 1.14 is infra-red (IR) spectrum of Homoharringtonine
salicyclate in the solid state. FIG. 1.14 (i) is IR(ATR) spectrum
in the crystalline state. FIG. 1.14 (ii) is IR(ATR) spectrum in the
amorphous state (film).
[0056] FIG. 2.2.1 is single crystal x-ray diffraction of
homoharringtonine base, form A (ORTEP-3 software).
[0057] FIG. 2.2.2 is single crystal x-ray diffraction of
homoharringtonine base, form B (ORTEP-3 software).
[0058] FIG. 2.2.3 is single crystal x-ray diffraction of
homoharringtonine base, form B with corresponding packing with unit
cell content (PLUTO drawing, ORTEP-3 software).
[0059] FIG. 2.3.1 is single crystal x-ray diffraction of
homoharringtonine hydrogen (2S)-(-)-malate (ORTEP-3 software).
[0060] FIG. 2.3.2 is single crystal x-ray diffraction of
homoharringtonine hydrogen (2S)-(-)-malate with corresponding
packing with unit cell content (PLUTO drawing, ORTEP-3
software).
[0061] FIG. 2.3.3 is x-ray powder diffraction (XRPD) of
homoharringtonine hydrogen (2S)-(-)-malate.
[0062] FIG. 2.4.1 is single crystal X-ray diffraction of
homoharringtonine hydrogen (2R)-(+)-malate (created by ORTEP-3
software).
[0063] FIG. 2.4.2 is single crystal X-ray diffraction of
homoharringtonine hydrogen (2R)-(+)-malate with corresponding
packing with unit cell content (PLUTO drawing, ORTEP-3
software).
[0064] FIG. 2.4.3 is X-ray powder diffraction (XRPD) of
homoharringtonine hydrogen (2R)-(+)-malate.
[0065] FIG. 2.5.1 is single crystal X-ray diffraction of
homoharringtonine hydrogen (2S,3S)-(-)-tartarate (created by
ORTEP-3 software).
[0066] FIG. 2.5.2 is single crystal X-ray diffraction of
homoharringtonine hydrogen (2S,3S)-(+)-tartarate with corresponding
packing with unit cell content (PLUTO drawing, ORTEP-3
software).
[0067] FIG. 2.5.3 is X-ray powder diffraction (XRPD) of
homoharringtonine hydrogen (2S,3S)-(-)-tartarate.
[0068] FIG. 2.6.1 is single crystal X-ray diffraction of
homoharringtonine hydrogen (2R,3R)-(+)-tartarate (created by
ORTEP-3 software).
[0069] FIG. 2.6.2 is single crystal X-ray diffraction of
homoharringtonine hydrogen (2R,3R)-(+)-tartarate with corresponding
packing with unit cell content (PLUTO drawing, ORTEP-3
software).
[0070] FIG. 2.6.3 is X-ray powder diffraction (XRPD) of
homoharringtonine hydrogen (2R,3R)-(+)-tartarate.
[0071] FIG. 2.7.1 is X-ray powder diffraction (XRPD) of
homoharringtonine hydrogen (2'''S)-citramalate.
[0072] FIG. 2.8.1 is single crystal X-ray diffraction of
homoharringtonine hydrogen (2R)-(-)-citramalate (created by ORTEP-3
software).
[0073] FIG. 2.8.2 is single crystal X-ray diffraction of
homoharringtonine hydrogen (2R)-(-)-citramalate with corresponding
packing with unit cell content (PLUTO drawing, ORTEP-3
software).
[0074] FIG. 2.8.3 is X-ray powder diffraction (XRPD) of
homoharringtonine hydrogen (2R)-(-)-citramalate.
[0075] FIG. 2.9.1 is single crystal X-ray diffraction of
homoharringtonine hydrogen itaconate (created by ORTEP-3
software).
[0076] FIG. 2.9.2 is single crystal X-ray diffraction of
homoharringtonine hydrogen itaconate with corresponding packing
with unit cell content (PLUTO drawing, ORTEP-3 software).
[0077] FIG. 2.10.1 is X-ray powder diffraction (XRPD) of
homoharringtonine hydrogen fumarate.
[0078] FIG. 2.11.1 is single crystal X-ray diffraction of
homoharringtonine dihydrogen citrate (created by ORTEP-3
software).
[0079] FIG. 2.11.2 is single crystal X-ray diffraction of
homoharringtonine dihydrogen citrate with corresponding packing
with unit cell content (PLUTO drawing, ORTEP-3 software).
[0080] FIG. 2.11.3 is X-ray powder diffraction (XRPD) of
homoharringtonine dihydrogen citrate.
[0081] FIG. 2.12.1 is single crystal X-ray diffraction of
homoharringtonine salicyclate (created by ORTEP-3 software).
[0082] FIG. 2.12.2 is single crystal X-ray diffraction of
homoharringtonine salicyclate (PLUTO drawing).
[0083] FIG. 2.12.3 is single crystal X-ray diffraction of
homoharringtonine salicyclate (stick drawing).
[0084] FIG. 2.12.4 is single crystal X-ray diffraction of
homoharringtonine salicydate with corresponding packing with unit
cell content (PLUTO drawing, ORTEP-3 software).
[0085] FIG. 3.1 is DSC pattern of homoharringtonine base.
[0086] FIG. 3.2 is DSC pattern of homoharringtonine hydrogen
(2S)-malate.
[0087] FIG. 3.3 is DSC pattern of homoharringtonine hydrogen
(2R)-malate.
[0088] FIG. 3.4 is DSC pattern of homoharringtonine hydrogen
(2S,3S)-tartrate.
[0089] FIG. 3.5 is DSC pattern of homoharringtonine hydrogen
(2R,3R)-tartrate.
[0090] FIG. 3.6 is DSC pattern of homoharringtonine hydrogen
(2S)-citramalate.
[0091] FIG. 3.7 is DSC pattern of homoharringtonine hydrogen
(2R)-citramalate.
[0092] FIG. 3.8 is DSC pattern of homoharringtonine hydrogen
succinate.
[0093] FIG. 3.9 is DSC pattern of homoharringtonine hydrogen
itaconate.
[0094] FIG. 3.10 is DSC pattern of homoharringtonine hydrogen
fumarate.
[0095] FIG. 3.11 is DSC pattern of homoharringtonine hydrogen
tartronate.
[0096] FIG. 3.12 is DSC pattern of homoharringtonine hydrogen
malonate.
[0097] FIG. 3.13 is DSC pattern of homoharringtonine dihydrogen
citrate.
[0098] FIG. 3.14 is DSC pattern of homoharringtonine
salicyclate.
DETAILED DESCRIPTION
[0099] In one embodiment, the crystalline salts of the invention
are used as drug substance for blending alone or in combination
with other therapeutical agents in pharmaceutical composition
useful as immunomodulating agents, particularly in using oral or
parenteral modes of administration.
[0100] A major embodiment of the invention is a new efficient
process of purification of natural, semi-synthetic or synthetic
version of harringtonines and their analogs using formation of a
crystallogenic salt and its fractional crystallization in organic
solvents, all the resulting purified compounds having the same
level of purity.
[0101] Another aspect of the invention is a new efficient process
of purification of natural homoharringtonine using formation of
crystallogenic salts and their fractional crystallization in
organic solvents giving the same level of purity as
homoharringtonine of hemi/semi-synthetic origin.
[0102] Another aspect of the invention is a new efficient process
of purification of natural harringtonine using formation of
crystallogenic salts and their fractional crystallization in
organic solvents giving the same level of purity as harringtonine
of hemi/semi-synthetic origin.
[0103] In one embodiment, the present invention relates to a
harringtonines salt in the crystalline state exhibiting a
protonated nitrogen seen in solid state analysis and having formula
1,
##STR00006##
[0104] comprising solvate, made by reacting a cephalotaxine ester
having formula 2,
##STR00007##
[0105] in which R.sup.1 is, but not limited to, alkyl, aryl,
cycloalkyl, heteroalkyl, heteroaryl or heterocycloalkyl, and
R.sup.2 is, independently, but not limited to H, alkyl, aryl,
cycloalkyl, heteroalkyl, heteroaryl or heterocydoalkyl, with an
acid having general formula AH in a crystallization solvent,
wherein the said salt has a water or alkohol solubility ranged
approximately from 5 mg/mL to approximately 100 mg/Ml.
[0106] In a preferred embodiment, the acid having general formula
AH is an organic acid. In a preferred embodiment, the organic acid
is selected among the following list: fumaric, maleic, citramalic,
malic, tartaric, tartronic, succinic, itaconic, citric acid or
salicylic acid.
[0107] A preferred embodiment of the invention is a crystalline
homohaningtonine hydrogen 2S-malate having substantially the same
IR spectrum, in the solid state as set out in FIG. 1.2, the same
single crystal X-ray diffractogram as set out in FIGS. 2.3.1 and
2.3.2, the same X-ray powder pattern as set out in FIG. 2.3.3 and
the same DSC curve as set out in FIG. 3.2.
[0108] A further preferred embodiment of the invention provides a
crystalline homoharringtonine hydrogen 2R-malate having
substantially the same IR spectrum, in the solid state as set out
in FIG. 1.3, the same single crystal X-ray diffractogram as set out
in FIGS. 2.4.1 and 2.4.2, the same X-ray powder pattern as set out
in FIG. 2.4.3 and the same DSC curve as set out in FIG. 3.3.
[0109] A further preferred aspect of the invention is a crystalline
homoharringtonine hydrogen (2S,3S)-tartrate having substantially
the same IR spectrum, in the solid state as set out in FIG. 1.4,
the same single crystal X-ray diffractogram as set out in FIGS.
2.5.1 and 2.5.2, the same X-ray powder pattern as set out in FIG.
2.5.3 and the same DSC curve as set out in FIG. 3.4.
[0110] Yet, a further embodiment of the invention is a crystalline
homoharringtonine hydrogen (2R,3R)-tartrate having substantially
the same IR spectrum, in the solid state as set out in FIG. 1.5,
the same single crystal X-ray diffractogram as set out in FIGS.
2.6.1 and 2.6.2, the same X-ray powder pattern as set out in FIG.
2.6.3 and the same DSC curve as set out in FIG. 3.5.
[0111] Yet, another embodiment of the invention provides a
crystalline homoharringtonine hydrogen (2S)-citramalate having
substantially the same IR spectrum, in the solid state as set out
in FIG. 1.6, the same X-ray powder pattern as set out in FIG. 2.7.1
and the same DSC curve as set out in FIG. 3.6.
[0112] Yet, a preferred aspect of this invention is a crystalline
homoharringtonine hydrogen (2R)-citramalate having substantially
the same IR spectrum, in the solid state as set out in FIG. 1.7,
the same single crystal X-ray diffractogram as set out in FIGS.
2.8.1 and 2.8.2, the same X-ray powder pattern as set out in FIG.
2.8.3 and the same DSC curve as set out in FIG. 3.7.
[0113] Yet, another preferred aspect of this invention provides a
crystalline homoharringtonine hydrogen succinate having
substantially the same IR spectrum, in the solid state as set out
in FIG. 1.8, and the same DSC curve as set out in FIG. 3.8.
[0114] Yet, a further preferred aspect of this invention is a
crystalline homoharringtonine hydrogen itaconate having
substantially the same IR spectrum, in the solid state as set out
in FIG. 1.9, the same single crystal X-ray diffractogram as set out
in FIGS. 2.9.1 and 2.9.2 and the same DSC curve as set out in FIG.
3.9.
[0115] Yet, a preferred aspect of this invention provides a
crystalline homoharringtonine hydrogen fumarate having
substantially the same IR spectrum, in the solid state as set out
in FIG. 1.10, the same X-ray powder pattern as set out in FIG.
2.10.1 and the same DSC curve as set out in FIG. 3.10.
[0116] Yet, an another aspect of the invention provides a
crystalline homohamrngtonine hydrogen tartronate having
substantially the same IR spectrum, in the solid state as set out
in FIG. 1.11 and the same DSC curve as set out in FIG. 3.11.
[0117] In addition, another embodiment provides a crystalline
homoharringtonine hydrogen malonate having substantially the same
IR spectrum, in the solid state as set out in FIG. 1.12 and the
same DSC curve as set out in FIG. 3.12.
[0118] Moreover, a preferred embodiment of this invention provides
a crystalline homoharringtonine dihydrogen citrate having
substantially the same IR spectrum, in the solid state as set out
in FIG. 1.13, the same single crystal X-ray diffractogram as set
out in FIGS. 2.11.1 and 2.11.2, the same X-ray powder pattern as
set out in FIG. 2.11.3 and the same DSC curve as set out in FIG.
3.13.
[0119] Also, a preferred aspect of this invention provides a
crystalline homoharringtonine hydrogen salicylate having
substantially the same IR spectrum, in the solid state as set out
in FIG. 1.14, the same single crystal X-ray diffractogram as set
out in FIGS. 2.12.1, 2.12.2, 2.12.3 and 2.12.4 and the same DSC
curve as set out in FIG. 3.14.
[0120] Yet, a preferred aspect of this invention provides a
pharmaceutical composition comprising an effective amount of one of
the salts of this invention, together with one or more
pharmaceutical acceptable inactive components such as carriers,
excipients, adjuvants or diluents.
[0121] Yet, a preferred aspect of this invention provides a
pharmaceutical dosage form dedicated to an oral mode of
administration selected among, for example, capsules, dragees,
emulsions, granules, pills, powders, solutions, suspensions,
tablets, microemulsions, elixirs, syrups, tea or powders for
reconstitution.
[0122] Yet, an another aspect of this invention provides a
pharmaceutical dosage form dedicated to a subcutaneous mode of
administration in non-acidic condition allowing a good locale
tolerance.
[0123] Another aspect of the invention is the use of at least the
solid form of one salt described in the invention for preparing the
above pharmaceutical composition as (i) chemotherapeutic agent,
(ii) enhancer of other chemotherapeutic agents (iii) after failure
of other agents (iv) for inhibiting tumors growth in animal, (v)
for inhibiting mammalian parasites, (vi) as immunosuppressive
agent, or (vii) as reversal agent.
[0124] A preferred embodiment of the invention describes a method
for treating mammalian tumors which comprises oral administering to
a mammal an antitumor effective amount of the solid form of one
salt described in this invention.
[0125] A further preferred embodiment of the invention describes a
method for treating mammalian tumors which comprises implantable
pharmaceutical preparation administering to a mammal an antitumor
effective amount of the solid form of at least one salt described
in this invention.
[0126] Yet, invention is also concerned with the use of solid form
of the salts of the invention as defined above, for the preparation
of pharmaceutical compositions for the treatment of cancer,
particularly brain cancer such as for example, but not limited to,
neuroblastoma and eventually their metastasis, lung cancer such as
for example non-small cells lung carcinoma eventually their
metastasis, ovarian high-grade carcinoma, breast cancer including
triple negative breast carcinoma and eventually their metastasis,
and pancreatic cancer including ductal adenocarcinoma, this therapy
being given alone or combined with at least another
chemotherapeutic agent, eventually combined with radiotherapy.
[0127] Another embodiment of the present invention relates to a
method of treating cancer, particularly brain cancer such as for
example, but not limited to, neuroblastoma and eventually their
metastasis, and lung cancer such as for example non-small cells
lung carcinoma eventually their metastasis, ovarian high-grade
carcinoma, breast cancer including triple negative breast carcinoma
and eventually their metastasis, and pancreatic cancer including
ductal adenocarcinoma, comprising administering to a patient or an
animal in need thereof a pharmaceutical composition comprising
solid form of the salts of the invention, said pharmaceutical
composition being administered alone or combined with at least
another chemotherapeutic agent, eventually combined with
radiotherapy.
[0128] Furthermore, invention also deals with the use of solid form
of the salts of the invention as defined above, for the preparation
of pharmaceutical compositions for the treatment of autoimmune
diseases, such as for example but not limited to multiple
sclerosis, psoriasis, rheumatoid arthritis, dermatomyositis,
Hashimoto's thyroiditis, systemic lupus erythematosus, this therapy
being given alone or combined with at least another
chemotherapeutic agent, said at least another chemotherapeutic
agent being eventually combined with radiotherapy, or with at least
another immunomodulating agent. Another embodiment of the present
invention relates to a method of treating autoimmune diseases, such
as for example but not limited to multiple sclerosis, psoriasis,
rheumatoid arthritis, dermatomyositis, Hashimoto's thyroiditis,
systemic lupus erythematosus, comprising administering to a patient
or an animal in need thereof a pharmaceutical composition
comprising solid form of the salts of the invention, said
pharmaceutical composition being administered alone or combined
with at least another chemotherapeutic agent, said at least another
chemotherapeutic agent being eventually combined with
radiotherapy.
[0129] Finally, the invention is also concerned with the use of
solid form of the salts of the invention as defined above, for the
preparation of pharmaceutical compositions for the treatment of
leukemias particularly acute myelod leukemia (AML), myelodysplastic
syndrome (MDS) and myeloproliferative disorders including chronic
myelogenous leukemia, polycythemia vera, essential thrombocythemia,
myelosclerosis, and/or lymphoma such as but not limited to a
multiple myeloma, a Hodgkin disease or a Burkitt lymphoma, said
therapy being given alone or combined with at least another
chemotherapeutic agent and/or with radiotherapy.
[0130] Another embodiment of the present invention relates to a
method of treating leukemias particularly acute myelod leukemia
(AML), myelodysplastic syndrome (MDS) and myeloproliferative
disorders including chronic myelogenous leukemia, polycythemia
vera, essential thrombocythemia, myelosclerosis, and/or, said
method comprising administering to a patient or an animal in need
thereof a pharmaceutical composition comprising solid form of the
salts of the invention, said pharmaceutical composition being
administered alone or combined with at least another
chemotherapeutic agent, including the targeted one, eventually
combined with radiotherapy.
[0131] In a preferred embodiment, the patient in need thereof
according to the present invention is a de novo patient or a
patient for which other therapy as failed to treat the disease from
which he suffers.
Example 1: General Procedure for Experimental Methods
[0132] 1.1 General Procedures for Salts Preparation
[0133] Cation and anion components are dissolved separately in a
solvent at a concentration close of saturation and at a temperature
close of boiling then both solutions are mixed under stirring then
slowly cooled and evaporated. After a period ranging from a few
minutes up to several days, crystal salt is collected. A sample of
the batch of crystals is kept suspended in its mother liquors for
the subsequent X-ray diffraction analysis. The remainder of the
batch was dried under vacuum for further solid characterization,
comparative stability studies and drug formulation.
[0134] 1.2 General Procedures for Solid State Characterization
[0135] Single Crystal X-Ray Diffractions Material and Methods
[0136] KappaCCD, Nonius diffractometer, Mo--K.sub..alpha. radiation
(.lamda.=0.71073 .ANG.). The structure was solved by direct methods
using the SHELXS-97 program [Sheldrick G. M., Acta Cryst. A64
(2008), 112-122], and then refined with full-matrix least-square
methods based on F.sup.2 (SHELXL-2013) [Sheldrick G. M., (2013)]
with the aid of the WINGX [L. J. Farrugia, J. Appl. Cryst., 2012,
45, 849-854] program. All non-hydrogen atoms were refined with
anisotropic atomic displacement parameters. Except nitrogen and
oxygen linked hydrogen atoms that were introduced in the structural
model through Fourier difference maps analysis, H atoms were
finally included in their calculated positions.
[0137] Collected information: atomic positions; unit cell
composition; crystal packing anisotropic displacement parameters;
bond lengths, dihedral and torsion angles, hydrogen bounding.
[0138] Original files with all parameters are includes on a CD and
may be visualized and handled in using ORTEP-3 software (ORTEP=Oak
Ridge Thermal-Ellipsoid Plot Program) available free of charge on
the Internet:
[0139] http://www.chem.gla.ac.uk/.about.louis/software/ortep3/
[0140] X-Ray Diffraction Powder
[0141] Diagrams were measured on a Bruker AXS D8 Advance
diffractometer, Bragg-Brentano geometry (.theta.-2 .theta.), CuK
.alpha.=1.5406 .ANG., 600 ms/pixel, rotation: 0.25/sec. For each
chart, the calculated pattern from the single crystal structure,
when available, is upped mentioned.
[0142] Differential Scanning Calorimetry (DSC)
[0143] The DSC analysis was performed using a Perkin Elmer DSC 4000
apparatus. The scan rate was 5.degree. C./min and the scanning
range of of temperature 40 to 230.degree. C. The accurately weighed
quantity was ranged from 1 to 3 mg. All operations were performed
under nitrogen atmosphere. The measured values were the Onset, the
Peak and the value of the free enthalpy variation. The eventual
product decomposition and the vaporization of solvent
crystallization (methanol and/or water) were recorded. The value of
the change in free energy, was given only as a guideline to assess
the endothermicity or exotermicity of the transition.
[0144] Melting Point Checking
[0145] Melting points were measured manually for visual checking of
the one determined with DSC. A Bucchi B-545 melting point apparatus
was used and mp are uncorrected.
[0146] Infrared Spectra
[0147] All vibrational spectra were recorded on a Perkin Elmer IR
FT Spectrum 2 apparatus equipped with diamond ATR accessory that is
to say using Attenuated Total Reflection technique. The crystalline
solids were crushed directly by in situ compression on the diamond
window and the amorphous state has been demonstrated by dissolving
the product in deuterated methanol then generating the film by in
situ evaporation on the diamond window.
[0148] 1.3 General Procedures for Liquid State and Solution
Characterizations
[0149] Nuclear Magnetic Resonance
[0150] NMR spectra were recorded automatically on a Bruker Avance
III spectrometer NanoBay--400 MHz (9.4 Tesla magnet) with a
BBFO+probe and sampler 120 positions, allows for automatic mode NMR
experiments one and two dimensions mainly for nuclei: 1H, 2H, 11B,
13C, 15N, 19F, 27Al, 31P, 119Sn or on Bruker Avance III--600 MHz
spectrometer.
[0151] Dissolving salts for .sup.13C NMR: 30 mg of compound were
dissolved in 600 .mu.L (5% m/V) of methanol D.sub.4 or deuterium
oxyde (or both if specified)
[0152] Water suppression: The irradiation technique known as
`watergate` (Selective pulse flanked by gradient pulses) was used
for proton NMR in the presence of D.sub.2O and/or MOD.sub.4 as
solvents.
[0153] High Performance Liquid Chromatography
[0154] Routine experiments were performed on a Waters HPLC-MS-DAD
coupled system (3100 pump, DAD 996 detector, 3100 mass
detector).
[0155] Solubility Determination
[0156] Solubility in water at 25.degree. C. was measured
semi-quantitatively at a threshold of 5 g per 100 mL. All the
homoharringtonine salts described in the below examples, unless
otherwise stated, are soluble at this threshold. Homoharringtonine
base itself is soluble at a threshold mower than 0.1 g per mL
Example 2: Analyses of Homoharringtonine Base for Comparison with
its Salts
##STR00008##
[0158] 2.1 Analysis of Homoharringtonine Base Alkaloid
[0159] Commercial homoharringtonine is provided by Sigma
Aldrich.NMR spectra were performed in deuterated methanol for
comparison with salt in the same solvent
[0160] By methanol recrystallisation of a commercial alkaloid from
natural source, it results fine white prisms (mp 145-146.degree.,
by DSC, see FIG. 3.1) used for all experiences.
[0161] .sup.1H NMR (400 MHz, Benzene-d.sub.6) .delta. 6.54 (s, 1H),
6.46 (s, 1H), 6.21-6.12 (m, 1H), 5.47 (d, J=1.4 Hz, 1H), 5.33 (d,
J=1.4 Hz, 1H), 4.67 (s, 1H), 3.43 (d, J=9.8 Hz, 1H), 3.34 (s, 3H),
3.28 (s, 3H), 2.83 (td, J=8.5, 4.5 Hz, 1H), 2.75 (dd, J=11.5, 4.5
Hz, 1H), 2.55 (dd, J=10.8, 7.5 Hz, 1H), 2.41 (dd, J=16.2, 6.9 Hz,
2H), 2.23-2.11 (m, 2H), 1.78 (m, 1H), 1.67-1.56 (m, 2H), 1.48 (m,
5H), 1.34-1.19 (m, 2H), 1.04 (d, J=6.7 Hz, 6H).
[0162] .sup.1H NMR (300 MHz, Chloroform-d) .delta. 6.62 (s, 1H),
6.54 (s, 1H), 6.00 (d, J=9.8 Hz, 1H), 5.87 (s, 2H), 5.05 (s, 1H),
3.78 (d, J=9.8 Hz, 1H), 3.68 (s, 3H), 3.57 (s, 3H), 3.52 (s, 1H),
3.20-3.04 (m, 2H), 3.01-2.88 (m, 1H), 2.60 (t, J=7.2 Hz, 1H), 2.38
(dd, J=13.7, 6.3 Hz, 1H), 2.26 (d, J=16.5 Hz, 1H), 2.10-1.97 (m,
1H), 1.91 (d, J=16.5 Hz, 1H), 1.75 (s, OH), 1.39 (dd, J=13.5, 6.4
Hz, 5H), 1.19 (s, 7H).
[0163] .sup.1H NMR (400 MHz, Methanol-d.sub.4)*.delta. 6.7 (s, 1H),
6.59 (s, 1H), 5.98 (dd, J=9.8, 0.8 Hz, 1H), 5.89 (d, J=1.2 Hz, 1H),
5.85 (d, J=1.2 Hz, 1H), 5.22 (d, J=0.8 Hz, 1H), 3.89 (d, J=9.8 Hz,
1H), 3.70 (s, 3H), 3.55 (s, 3H), 3.20 (ddd, J=14.1, 12.4, 7.9 Hz,
1H), 2.96 (m, 1H), 2.88 (m, 1H), 2.64 (dd, J=11.4, 7.6 Hz, 1H),
2.44 (dd, J=14.3, 6.8 Hz, 1H), 2.17 (d, J=16.1 Hz, 1H), 2.03 (m,
1H), 1.95 (m, 1H), 1.90 (d, J=16.1 Hz, 1H), 1.49-1.30 (m, 5H), 1.25
(dd, J=9.8, 5.8 Hz, 1H), 1.17 (s, 3H), 1.16 (s, 3H). *Partial
presuppression of water signal using `watergate` irradiation
[0164] .sup.13C NMR (101 MHz, MeOD) .delta. 174.68, 171.76, 159.97,
148.21, 147.32, 134.49, 129.88, 114.02, 110.86, 102.10, 100.74,
76.03, 75.52, 72.14, 71.35, 58.04, 56.48, 54.60, 52.00, 49.64,
44.89, 44.15, 43.86, 40.87, 32.08, 29.27, 29.01, 20.82, 19.19.
[0165] IR (KBr, solid), cm.sup.-1 3551.9, 3412.3, 3000.4, 2976.1,
2966.0, 2958.6, 2911.4, 2876.0, 2814.4, 2740.8, 1743.0, 1653.5,
1624.7, 1505.3, 1488.1, 1454.8, 1436.1, 1411.2, 1392.8, 1377.7,
1367.2, 1346.3, 1306.4, 1274.3, 1261.5, 1230.0, 1190.8, 1162.1,
1135.3, 1119.9, 1082.0, 1027.9, 1000.5, 932.1, 900.6, 879.3, 854.2,
827.3, 804.9, 795.2, 772.4, 762.9, 738.3, 705.7, 674.0, 661.4,
610.8, 556.7, 540.9, 522.1, 512.8, 503.3. See FIG. 1.1
[0166] A) Single Crystal X Ray Diffraction of Homoharringtonine
Base (Form A).
[0167] See Corresponding FIG. 2.2.1
[0168] From a suspension in its mother liquor, a suitable single
crystal of size 0.5.times.0.4.times.0.4 mm was finally selected and
implemented on the diffractometer.
TABLE-US-00002 Structural data: Formula weight 545.61 Temperature
293(2)K Wavelength 0.71073 .ANG. Crystal system, space group
orthorhombic, P 2.sub.1 2.sub.1 2.sub.1 Unit cell dimensions a =
11.9512(2) .ANG., .alpha. = 90.degree. b = 15.2211(2) .ANG., .beta.
= 90.degree. c = 15.9670(2) .ANG., .gamma. = 90.degree. Volume
2904.56(7) .ANG..sup.3 Z, Calculated density 4, 1.248 (g cm.sup.-1)
Absorption coefficient 0.092 mm.sup.-1 F(000) 1168 Crystal size 0.5
.times. 0.4 .times. 0.4 mm Crystal color colourless Theta range for
data collection 2.881 to 29.046.degree. h_min, h_max -16, 16 k_min,
k_max -20, 20 l_min, l_max -21, 21 Reflections collected/unique
35627/7642 [.sup.aR(int) = 0.049] Reflections [I > 2.sigma.]
5925 Completeness to theta_max 0.99 Absorption correction type none
Refinement method Full-matrix least-squares on F.sup.2
Data/restraints/parameters 7642/0/352 .sup.bGoodness-of-fit 1.034
Final R indices [I > 2.sigma.] .sup.cR.sub.1 = 0.0495,
.sup.dwR.sub.2 = 0.1256 R indices (all data) .sup.cR.sub.1 =
0.0719, .sup.dwR.sub.2 = 0.1411 Largest diff. peak and hole 0.284
and -0.203 e..ANG..sup.-3
Atomic coordinates, site occupancy (%) and equivalent isotropic
displacement parameters (A.sup.2.times.10.sup.3). U(eq) is defined
as one third of the trace of the orthogonalized Uij tensor. Atom
numbering of FIG. 2.2.1 corresponds to below table.
TABLE-US-00003 Atom x y z occ. U(eq) C1 0.9387(2) 0.25439(15)
0.89639(14) 1 0.0374(5) H1 1.0075 0.2226 0.8823 1 0.045 C2
0.9724(2) 0.34995(16) 0.90497(14) 1 0.0390(5) C3 1.0805(2)
0.37308(17) 0.87926(17) 1 0.0442(5) H3 1.1276 0.3319 0.8546 1 0.053
C4 1.1150(2) 0.45761(19) 0.89130(17) 1 0.0494(6) C5 1.2096(4)
0.5829(2) 0.8960(3) 1 0.0862(12) H5A 1.2163 0.6192 0.8463 1 0.103
H5B 1.2695 0.5986 0.9342 1 0.103 C6 1.0469(3) 0.51945(17)
0.92762(18) 1 0.0528(6) C7 0.9399(2) 0.49967(18) 0.95274(17) 1
0.0498(6) H7 0.8939 0.5421 0.9765 1 0.06 C8 0.9026(2) 0.41337(17)
0.94127(15) 1 0.0434(5) C9 0.7884(2) 0.38828(18) 0.97226(17) 1
0.0493(6) H9A 0.7497 0.3553 0.9291 1 0.059 H9B 0.7455 0.4411 0.9836
1 0.059 C10 0.7951(3) 0.3328(2) 1.05191(18) 1 0.0569(7) H10A 0.8046
0.3714 1.0997 1 0.068 H10B 0.7251 0.3014 1.0592 1 0.068 C11
0.8990(3) 0.2199(3) 1.12687(17) 1 0.0648(8) H11A 0.8277 0.1965
1.1455 1 0.078 H11B 0.9302 0.256 1.1711 1 0.078 C12 0.9767(5)
0.1482(3) 1.1039(2) 1 0.1012(16) H12A 1.0519 0.162 1.1227 1 0.121
H12B 0.9536 0.0937 1.1303 1 0.121 C13 0.9745(3) 0.1391(2) 1.0129(2)
1 0.0677(9) H13A 0.949 0.0808 0.9975 1 0.081 H13B 1.0488 0.148 0.99
1 0.081 C14 0.8927(2) 0.20973(17) 0.97856(14) 1 0.0426(5) C15
0.7844(2) 0.16871(17) 0.95088(16) 1 0.0457(5) H15 0.7362 0.1387
0.9865 1 0.055 C16 0.7655(2) 0.18031(16) 0.86960(15) 1 0.0407(5)
C17 0.8541(2) 0.23162(15) 0.82622(14) 1 0.0374(5) H17 0.8905 0.1948
0.7839 1 0.045 C18 0.8201(2) 0.31714(16) 0.70344(14) 1 0.0415(5)
C19 0.7614(2) 0.39953(17) 0.67082(15) 1 0.0470(6) C20 0.7996(3)
0.48109(18) 0.71967(18) 1 0.0574(7) H20A 0.7501 0.5295 0.7059 1
0.069 H20B 0.7916 0.4693 0.7791 1 0.069 C21 0.9168(4) 0.5087(2)
0.7031(2) 1 0.0712(10) C22 1.1000(4) 0.4595(4) 0.6716(3) 1
0.1074(16) H22A 1.1041 0.4831 0.6159 1 0.161 H22B 1.1425 0.4061
0.6745 1 0.161 H22C 1.1301 0.5014 0.7106 1 0.161 C23 0.6346(2)
0.38782(19) 0.67748(17) 1 0.0525(6) H23A 0.6135 0.3894 0.7361 1
0.063 H23B 0.5983 0.4369 0.6499 1 0.063 C24 0.5914(3) 0.3023(2)
0.6389(2) 1 0.0590(7) H24A 0.6233 0.2952 0.5834 1 0.071 H24B 0.6159
0.2531 0.6729 1 0.071 C25 0.4642(3) 0.3012(2) 0.6326(2) 1 0.0665(8)
H25A 0.4338 0.3092 0.6884 1 0.08 H25B 0.4411 0.3513 0.5994 1 0.08
C26 0.4115(3) 0.2193(3) 0.5950(2) 1 0.0754(10) C27 0.2855(4)
0.2319(4) 0.5904(4) 1 0.1200(19) H27A 0.2691 0.2858 0.5617 1 0.18
H27B 0.2553 0.2343 0.6461 1 0.18
[0169] Single Crystal X Ray Diffraction of Homoharringtonine Base
(Form B)
[0170] See Corresponding FIGS. 2.2.2 and 2.2.3
[0171] From a suspension in its mother liquor, a suitable single
crystal of size 0.43.times.0.29.times.0.18 mm was finally selected
and implemented on the diffractometer.
TABLE-US-00004 Structural data Empirical formula C.sub.31H.sub.49N
O.sub.12 Extended formula C.sub.29H.sub.39N O.sub.9, 2(CH.sub.4O),
H.sub.2O Formula weight 627.71 Temperature 150(2)K Wavelength
0.71073 .ANG. Crystal system, space group orthorhombic, P 2.sub.1
2.sub.1 2.sub.1 Unit cell dimensions a = 11.7738(10) .ANG., .alpha.
= 90.degree. b = 14.3907(13) .ANG., .beta. = 90.degree. c =
19.1368(15) .ANG., .gamma. = 90.degree. Volume 3242.4(5)
.ANG..sup.3 Z, Calculated density 4, 1.286 (g cm.sup.-1) Absorption
coefficient 0.098 mm.sup.-1 F(000) 1352 Crystal size 0.43 .times.
0.29 .times. 0.18 mm Crystal color colourless Theta range for data
collection 3.02 to 27.46.degree. h_min, h_max -15, 13 k_min, k_max
-18, 18 l_min, l_max -24, 19 Reflections collected/unique
16236/4103 [.sup.aR(int) = 0.0334] Reflections [I > 2.sigma.]
3764 Completeness to theta_max 0.99 Absorption correction type
multi-scan Max. and min. transmission 0.982, 0.874 Refinement
method Full-matrix least-squares on F.sup.2
Data/restraints/parameters 4103/2/421 .sup.bGoodness-of-fit 1.031
Final R indices [I > 2.sigma.] .sup.cR.sub.1 = 0.0346,
.sup.dwR.sub.2 = 0.0871 R indices (all data) .sup.cR.sub.1 = 0.039,
.sup.dwR.sub.2 = 0.09 Largest diff. peak and hole 0.259 and -0.2
e..ANG..sup.-3
[0172] Atomic coordinates, site occupancy (%) and equivalent
isotropic displacement parameters (A.sup.2.times.10.sup.3).
[0173] U(eq) is defined as one third of the trace of the
orthogonalized Uij tensor.
[0174] Atom numbering of FIGS. 2.2.2 and 2.2.3 corresponds to below
table.
TABLE-US-00005 Atom x Y z occ. U(eq) C1 0.14461(16) 0.92123(13)
0.19855(10) 1 0.0185(4) H1 0.1645 0.9474 0.2424 1 0.022 C2
0.13507(16) 0.96935(13) 0.14015(10) 1 0.0182(4) C3 0.10057(15)
0.91236(13) 0.07788(10) 1 0.0162(4) H3 0.0245 0.9326 0.0604 1 0.019
C4 0.09461(15) 0.81122(13) 0.10658(10) 1 0.0156(4) H4 0.0135 0.7915
0.1024 1 0.019 C5 0.11993(15) 0.81891(13) 0.18697(10) 1 0.0173(4)
C6 0.01795(18) 0.78404(16) 0.22996(10) 1 0.0236(4) H6A -0.0435
0.831 0.2309 1 0.028 H6B -0.0124 0.7254 0.2103 1 0.028 C7
0.06647(19) 0.76830(18) 0.30351(12) 1 0.0324(5) H7A 0.0543 0.8236
0.3334 1 0.039 H7B 0.0307 0.7137 0.326 1 0.039 C8 0.19354(19)
0.75150(15) 0.29115(10) 1 0.0250(4) H8A 0.2394 0.8 0.3146 1 0.03
H8B 0.2164 0.69 0.3095 1 0.03 N9 0.21006(14) 0.75555(11) 0.21387(8)
1 0.0190(3) C10 0.32858(17) 0.78051(14) 0.19706(11) 1 0.0214(4)
H10A 0.3792 0.729 0.2115 1 0.026 H10B 0.3502 0.8363 0.2243 1 0.026
C11 0.34668(16) 0.80017(13) 0.11940(10) 1 0.0185(4) H11A 0.3221
0.8645 0.1092 1 0.022 H11B 0.4288 0.7958 0.1088 1 0.022 C12
0.28274(15) 0.73420(12) 0.07195(10) 1 0.0166(4) C13 0.16369(16)
0.74035(12) 0.06606(9) 1 0.0158(4) C14 0.10458(16) 0.68001(13)
0.02102(10) 1 0.0185(4) H14 0.0244 0.6838 0.0162 1 0.022 C15
0.16632(17) 0.61550(13) -0.01569(11) 1 0.0214(4) C16 0.28285(17)
0.60909(13) -0.00929(11) 1 0.0215(4) C17 0.34324(16) 0.66697(13)
0.03421(10) 1 0.0196(4) H17 0.4234 0.6616 0.0385 1 0.023 C18
0.2238(2) 0.49213(19) -0.07619(17) 1 0.0473(7) H18A 0.2304 0.4778
-0.1266 1 0.057 H18B 0.2144 0.433 -0.0504 1 0.057 C19 0.19032(19)
1.11428(14) 0.18498(12) 1 0.0283(5) H19A 0.2613 1.0871 0.2022 1
0.042 H19B 0.1334 1.1137 0.2224 1 0.042 H19C 0.2041 1.1785 0.1701 1
0.042 C21 0.15776(16) 0.96435(12) -0.03634(9) 1 0.0155(3) C22
0.25991(16) 0.96278(12) -0.08591(10) 1 0.0169(4) C23 0.29059(16)
0.86127(13) -0.10429(10) 1 0.0188(4) H23A 0.3594 0.8612 -0.134 1
0.023 H23B 0.3091 0.8276 -0.0607 1 0.023 C24 0.19701(16)
0.80987(13) -0.14192(10) 1 0.0187(4) C25 0.1561(2) 0.68927(17)
-0.22153(14) 1 0.0361(5) H25A 0.1117 0.7289 -0.2529 1 0.054 H25B
0.1053 0.6598 -0.1875 1 0.054 H25C 0.195 0.6412 -0.2488 1 0.054 C31
0.36248(16) 1.00778(13) -0.04883(10) 1 0.0195(4) H31A 0.3848 0.9679
-0.009 1 0.023 H31B 0.4272 1.0094 -0.0819 1 0.023 C32 0.34172(18)
1.10662(12) -0.02159(11) 1 0.0212(4) H32A 0.3272 1.149 -0.0613 1
0.025 H32B 0.2742 1.1072 0.0092 1 0.025 C33 0.44570(19) 1.13955(14)
0.01904(12) 1 0.0273(5) H33A 0.5137 1.1282 -0.0102 1 0.033 H33B
0.453 1.1004 0.0613 1 0.033 C34 0.44740(19) 1.24164(14) 0.04199(11)
1 0.0248(4) C35 0.5515(2) 1.25841(17) 0.08801(15) 1 0.0416(6) H35A
0.5496 1.2159 0.128 1 0.062 H35B 0.5509 1.3227 0.1048 1 0.062 H35C
0.6206 1.2474 0.0607 1 0.062 C36 0.4479(2) 1.30687(14) -0.01973(12)
1 0.0311(5) H36A 0.4469 1.3713 -0.0031 1 0.047 H36B 0.3805 1.2954
-0.0486 1 0.047 H36C 0.5164 1.2963 -0.0477 1 0.047 O1 0.14964(13)
1.06111(9) 0.12688(7) 1 0.0244(3) O2 0.12829(13) 0.55129(11)
-0.06415(9) 1 0.0329(4) O3 0.32361(13) 0.53951(11) -0.05252(9) 1
0.0328(4) O4 0.18522(11) 0.92032(9) 0.02297(7) 1 0.0169(3) O5
0.06782(11) 1.00004(10) -0.04854(7) 1 0.0215(3) O6 0.23721(13)
1.01397(10) -0.14731(7) 1 0.0221(3) HO6 0.170(3) 1.007(2)
-0.1577(16) 1 0.0 O7 0.09736(13) 0.82269(11) -0.13360(9) 1
0.0326(4) O8 0.23961(13) 0.74537(10) -0.18490(8) 1 0.0289(3) O9
0.34591(17) 1.25612(12) 0.08302(10) 1 0.0403(4) HO9 0.346(3)
1.315(2) 0.0956(15) 1 0.0 C51 0.1062(3) 0.5267(2) 0.2091(2) 1
0.0688(11 H51A 0.105 0.4613 0.1948 1 0.103 H51B 0.0453 0.5604
0.1851 1 0.103 H51C 0.0948 0.531 0.2597 1 0.103 O52 0.21050(17)
0.56578(12) 0.19155(11) 1 0.0451(5) H52 0.204(3) 0.632(3)
0.1905(18) 1 0.068 O61 0.03166(17) 1.03110(13) -0.21801(10) 1
0.0431(4) H61 -0.026(3) 1.028(3) -0.1858(18) 1 0.065 C62 -0.0021(2)
0.96665(19) -0.26999(14) 1 0.0428(6) H62A 0.0298 0.9852 -0.3152 1
0.064 H62B -0.0852 0.9655 -0.2731 1 0.064 H62C 0.0257 0.9046
-0.2576 1 0.064 O71 0.35707(17) 1.45528(12) 0.12174(10) 1 0.0408(4)
H71A 0.410(2) 1.489(2) 0.0914(15) 1 0.061 H71B 0.301(2) 1.498(2)
0.1443(16) 1 0.061
Example 3: Preparation and Analyses of Homoharringtonine Hydrogen
(2S)-Malate (Synonymous: Homoharringtonine (2S)-Bimalate)
##STR00009##
[0176] This ionic compound was obtained from commercial
homoharringtonine mixed with commercial (2S)-(-)-malic acid
(natural form) according to the general procedure in which the
solvent was methanol, then isolated as a white prismatic solid mp
205.4-207.7.degree. C. from MeOH (measured by DSC, see FIG. 3.2).
Several potentially acceptable crystals were kept suspended in
their mother liquors for the subsequent X-ray diffraction analysis.
(see below).
[0177] .sup.1H NMR (400 MHz. Methanol-d.sub.4)* .delta. 6.79 (s,
1H), 6.74 (s, 1H), 6.09 (dd, J=9.6, 0.6 Hz, 1H), 5.96 (d, J=1.1 Hz,
1H), 5.93 (d, J=1.1 Hz, 1H), 5.33 (d, J=0.6 Hz, 1H), 4.24 (dd,
J=7.4, 5.4 Hz, 1H), 4.16 (d, J=9.6 Hz, 1H), 3.81 (s, 3H), 3.54 (s,
3H), 3.50 (dd, J=9.5, 4.3 Hz, 1H), 3.42-3.32 (m, 1H), 3.21-3.10 (m,
1H), 2.76 (dd, J=15.9, 5.5 Hz, 1H), 2.71-2.62 (m, 1H), 2.48 (dd,
J=15.8, 7.4 Hz, 1H), 2.26-2.05 (m, 4H), 1.94 (d, J=16.1 Hz, 2H),
1.47-1.29 (m, 5H), 1.29-1.17 (m, 1H), 1.15 (s, 6H). *Partial
presuppression of water signal using `watergate` irradiation
[0178] .sup.1H NMR (600 MHz, Deuterium oxide)* .delta. 6.84 (s,
1H), 6.76 (s, 1H), 6.01 (dd, J=9.6, 0.7 Hz, 1H), 5.95 (d, J=1.0 Hz,
1H), 5.94 (d, J=1.0 Hz, 1H), 5.34 (d, J=0.6 Hz, 1H), 4.31 (dd,
J=8.2, 4.2 Hz, 1H), 4.19 (d, J=9.6 Hz, 1H), 3.76 (s, 3H), 3.52 (s,
3H), 3.52 (m, 1H), 3.42-3.32 (m, 1H), 3.30-3.23 (m, 1H), 3.22-3.15
(m, 1H), 2.76 (dd, J=16.0, 4.2 Hz, 1H), 2.74-2.68 (m, 1H), 2.57
(dd, J=16.0, 8.2 Hz, 1H), 2.36 (d, J=17.0 Hz, 1H), 2.29-2.08 (m,
2H), 1.99 s(d, J=16.9 Hz, 1H), 1.97-1.89 (m, 1H), 1.45-1.37 (m,
2H), 1.36-1.26 (m, 3H), 1.12 (s, 6H), 1.12-1.02 (m, 1H). *Partial
presuppression of water signal using `watergate` irradiation
[0179] .sup.13C NMR APT* (101 MHz, MeOD) .delta. 179.23, 176.13,
174.23, 171.61, 165.05, 149.76, 148.75, 130.92, 126.86, 114.85,
111.80, 102.86, 96.12, 78.09, 76.08, 74.35, 71.27, 69.35, 59.01,
54.21, 53.27, 52.07, 48.94, 44.76, 44.06, 41.80, 40.88, 40.52,
29.25, 29.23, 29.17, 19.95, 19.09.
[0180] .sup.13C NMR APT* (101 MHz, D.sub.2O) .delta. 178.97,
176.21, 174.23, 171.93, 162.88, 147.83, 146.74, 129.74, 125.22,
113.38, 111.12, 101.62, 95.52, 76.98, 75.25, 73.68, 71.34, 68.50,
58.41, 52.95, 52.24, 51.25, 47.58, 42.71, 42.54, 40.00, 39.18,
38.76, 27.69, 27.58, 27.47, 18.58, 17.68. *APT=Attached Proton
Test
[0181] IR (Diamond ATR, solid) cm-1 3404, 2969, 2601, 1981, 1758,
1736, 1712, 1657, 1525, 1505, 1490, 1468, 1435, 1374, 1353, 1265,
1226, 1188, 1148, 1080, 1032, 983, 943, 925, 862, 830, 796, 770,
756, 708, 691, 674, 650, 615, 589, 565, 541, 510, 477. See FIG.
1.2
[0182] IR (Diamond ATR, film) cm.sup.-1 3422, 2964, 1742, 1656,
1596, 1506, 1490, 1440, 1373, 1266, 1224, 1168, 1084, 1033, 929,
710, 615, 566, 509, 477, 0, 983, 943, 925, 862, 830, 796, 770, 756,
708, 691, 674, 650, 615, 589, 565, 541, 510. See FIG. 1.2
[0183] Solubility in neutral water higher than 60 mg/mL
[0184] A. Single Crystal X-Ray Diffraction (See FIGS. 2.3.1 and
2.3.2)
[0185] From a suspension in its mother liquor, a suitable single
crystal of size 0.58.times.0.46.times.0.29 mm was finally selected
and implemented on the diffractometer.
TABLE-US-00006 Structural data Empirical formula C.sub.33H.sub.45N
O.sub.14 Extended formula C.sub.29H.sub.40N O.sub.9,
C.sub.4H.sub.5O.sub.5 Formula weight 679.7 Temperature 150(2)K
Wavelength 0.71073 .ANG. Crystal system, space group orthorhombic,
P 2.sub.1 2.sub.1 2.sub.1 Unit cell dimensions a = 11.488(2) .ANG.,
.alpha. = 90.degree. b = 15.399(3) .ANG., .beta. = 90.degree. c =
18.825(4) .ANG., .gamma. = 90.degree. Volume 3330.2(11) .ANG..sup.3
Z, Calculated density 4, 1.356 (g cm.sup.-1) Absorption coefficient
0.106 mm.sup.-1 F(000) 1448 Crystal size 0.58 .times. 0.46 .times.
0.29 mm Crystal color white Theta range for data collection 3.09 to
27.48.degree. h_min, h_max -14, 14 k_min, k_max -19, 13 l_min,
l_max -18, 24 Reflections collected/unique 28567/4233 [.sup.aR(int)
= 0.1176] Reflections [I > 2.sigma.] 2414 Completeness to
theta_max 0.998 Absorption correction type multi-scan Max. and min.
transmission 0.970, 0.688 Refinement method: Full-matrix
least-squares on F.sup.2 Data/restraints/parameters 4233/0/440
.sup.bGoodness-of-fit 1.038 Final R indices [I > 2.sigma.]:
.sup.cR.sub.1 = 0.0735, .sup.dwR.sub.2 = 0.1727 R indices (all
data): .sup.cR.sub.1 = 0.1366, .sup.dwR.sub.2 = 0.2124 Largest
diff. peak and hole 0.555 and -0.27 e..ANG..sup.-3
Atomic coordinates, site occupancy (%) and equivalent isotropic
displacement parameters (A.sup.2.times.10.sup.3). U(eq) is defined
as one third of the trace of the orthogonalized Uij tensor.
[0186] Atom numbering of FIG. 2.3.1 corresponds to below table.
TABLE-US-00007 Atom x y z occ. U(eq) C1 0.8902(5) 0.0603(4)
0.7067(3) 1 0.0450(15) H1 0.8742 0.0363 0.7521 1 0.054 C2 0.8846(5)
0.0175(4) 0.6464(3) 1 0.0405(13) C3 0.9163(5) 0.0678(4) 0.5821(3) 1
0.0412(14) H3 0.9896 0.0445 0.5606 1 0.049 C4 0.9359(4) 0.1614(4)
0.6109(3) 1 0.0355(13) H4 1.0189 0.1762 0.6007 1 0.043 C5 0.9255(5)
0.1528(4) 0.6934(3) 1 0.0391(13) C6 1.0363(5) 0.1790(5) 0.7327(3) 1
0.0501(16) H6A 1.0975 0.1343 0.7269 1 0.06 H6B 1.0662 0.2352 0.7148
1 0.06 C7 0.9996(6) 0.1865(5) 0.8099(3) 1 0.0584(18) H7A 1.0501
0.2277 0.836 1 0.07 H7B 1.0023 0.1293 0.8339 1 0.07 C8 0.8744(6)
0.2204(5) 0.8049(3) 1 0.0539(17) H8A 0.8215 0.1843 0.834 1 0.065
H8B 0.8701 0.2812 0.8219 1 0.065 N9 0.8411(4) 0.2153(4) 0.7276(3) 1
0.0432(12) H9 0.8561 0.2696 0.7081 1 0.052 C10 0.7137(5) 0.1984(5)
0.7172(3) 1 0.0496(16) H10A 0.6691 0.2495 0.7338 1 0.06 H10B 0.6904
0.148 0.7467 1 0.06 C11 0.6826(4) 0.1802(4) 0.6402(3) 1 0.0436(14)
H11A 0.7034 0.1193 0.6287 1 0.052 H11B 0.5974 0.1864 0.6341 1 0.052
C12 0.7435(4) 0.2399(4) 0.5885(3) 1 0.0401(14) C13 0.8622(4)
0.2320(4) 0.5758(3) 1 0.0349(13) C14 0.9172(4) 0.2871(4) 0.5278(3)
1 0.0364(13) H14 0.9984 0.2823 0.5191 1 0.044 C15 0.8516(5)
0.3487(4) 0.4933(3) 1 0.0441(15) C16 0.7360(5) 0.3568(4) 0.5058(4)
1 0.0464(15) C17 0.6788(5) 0.3043(4) 0.5538(3) 1 0.0437(15) H17
0.598 0.3115 0.563 1 0.052 C18 0.7884(6) 0.4626(5) 0.4322(5) 1
0.076(2) H18A 0.8017 0.5203 0.4539 1 0.091 H18B 0.7749 0.4707
0.3807 1 0.091 C19 0.8154(7) -0.1118(5) 0.6969(4) 1 0.066(2) H19A
0.8787 -0.115 0.7317 1 0.099 H19B 0.7907 -0.1707 0.6841 1 0.099
H19C 0.7496 -0.0802 0.7175 1 0.099 C21 0.8355(4) 0.0154(4)
0.4745(3) 1 0.0382(13) C22 0.7306(5) 0.0234(4) 0.4248(3) 1
0.0412(14) C23 0.7103(5) 0.1202(4) 0.4059(3) 1 0.0379(13) H23A
0.6413 0.1246 0.3746 1 0.046 H23B 0.6932 0.1527 0.4501 1 0.046 C24
0.8129(5) 0.1618(4) 0.3693(3) 1 0.0409(14) C25 0.8695(6) 0.2717(5)
0.2887(4) 1 0.069(2) H25A 0.9088 0.31 0.3226 1 0.104 H25B 0.8353
0.3065 0.2505 1 0.104 H25C 0.9259 0.2308 0.2686 1 0.104 C31
0.6209(5) -0.0103(4) 0.4625(3) 1 0.0450(14) H31A 0.6041 0.028
0.5035 1 0.054 H31B 0.5544 -0.006 0.4293 1 0.054 C32 0.6293(5)
-0.1031(4) 0.4889(4) 1 0.0491(16) H32A 0.6941 -0.108 0.5233 1 0.059
H32B 0.6462 -0.1422 0.4484 1 0.059 C33 0.5166(6) -0.1309(5)
0.5242(4) 1 0.0596(19) H33A 0.4518 -0.1174 0.4914 1 0.072 H33B
0.5056 -0.0947 0.5672 1 0.072 C34 0.5053(7) -0.2260(5) 0.5462(4) 1
0.069(2) C35 0.6051(10) -0.2507(6) 0.5911(5) 1 0.100(3) H35A 0.5942
-0.3101 0.6086 1 0.15 H35B 0.6107 -0.2108 0.6315 1 0.15 H35C 0.6769
-0.2477 0.563 1 0.15 C36 0.3912(10) -0.2401(7) 0.5849(6) 1 0.112(4)
H36A 0.3267 -0.2205 0.5549 1 0.168 H36B 0.3914 -0.2069 0.6293 1
0.168 H36C 0.3818 -0.302 0.5955 1 0.168 O31 0.5016(6) -0.2752(4)
0.4798(3) 1 0.104(2) H31 0.4952 -0.3283 0.4889 1 0.156 O1 0.8553(4)
-0.0674(3) 0.6348(2) 1 0.0543(11) O2 0.6901(4) 0.4222(3) 0.4640(3)
1 0.0625(13) O3 0.8872(4) 0.4087(3) 0.4432(2) 1 0.0560(13) O21
0.8226(3) 0.0660(2) 0.53083(19) 1 0.0371(9) O22 0.9152(3)
-0.0342(3) 0.4646(2) 1 0.0507(11) O23 0.7478(3) -0.0256(3)
0.3627(2) 1 0.0426(10) H23 0.8145 -0.0155 0.3464 1 0.064 O24
0.9138(3) 0.1434(3) 0.3794(2) 1 0.0522(12) O25 0.7785(4) 0.2240(3)
0.3248(3) 1 0.0561(12) C51 0.8039(5) 0.4264(5) 0.7157(4) 1
0.0518(17) C52 0.7548(6) 0.5067(5) 0.6770(4) 1 0.0531(17) H52
0.7672 0.5593 0.7073 1 0.064 C53 0.6259(6) 0.4973(5) 0.6614(4) 1
0.060(2) H53A 0.606 0.5382 0.6226 1 0.072 H53B 0.6125 0.4379 0.6431
1 0.072 C54 0.5421(6) 0.5127(5) 0.7216(4) 1 0.065(2) O51 0.8885(4)
0.3887(3) 0.6897(3) 1 0.0557(12) O52 0.7540(6) 0.4053(4) 0.7716(3)
1 0.095(2) O53 0.8147(6) 0.5173(5) 0.6130(4) 1 0.100(2) H53 0.8805
0.4937 0.6161 1 0.15 O54 0.5697(6) 0.4866(5) 0.7821(3) 1 0.097(2)
H54 0.5978 0.4363 0.7791 1 0.146 O55 0.4515(5) 0.5523(5) 0.7122(3)
1 0.110(3)
[0187] B. X-Ray Powder Diffraction
[0188] The sample is pure and there is a very good match between
the experimental pattern and the calculated pattern (for view of
diagrams and experimental details, see FIG. 2.3.3)
Example 4: Preparation and Analyses of Homoharringtonine Hydrogen
(2R)-Malate (Diastereomer of Example 3)
##STR00010##
[0190] This ionic compound was obtained from commercial
homoharringtonine mixed with commercial (2R)-(+)-malic acid
(unnatural form) according to the general procedure in which the
solvent was methanol, then isolated as a white prismatic solid mp
205-208.degree. C. from MeOH (measured by DSC, see FIG. 3.3).
Several potentially acceptable crystals were kept suspended in
their mother liquors for the subsequent X-ray diffraction analysis.
(see below).
[0191] DSC Analysis (See FIG. 3.3)
[0192] .sup.1H NMR (400 MHz, Methanol-d.sub.4)*.delta. 6.80 (s,
1H), 6.74 (s, 1H), 6.09 (d, J=9.6 Hz, 1H), 5.97 (d, J=1.0 Hz, 1H),
5.93 (d, J=0.9 Hz, 1H), 5.33 (s, 1H), 4.26 (dd, J=7.4, 5.5 Hz, 1H),
4.17 (d, J=9.6 Hz, 1H), 3.81 (s, 3H), 3.55 (s, 3H), 3.53 (s, 1H),
3.34 (s, 2H), 3.22-3.12 (m, 1H), 2.77 (dd, J=15.9, 5.4 Hz, 1H),
2.72-2.64 (m, 1H), 2.49 (dd, J=15.9, 7.4 Hz, 1H), 2.29-2.05 (m,
4H), 1.95 (d, J=16.1 Hz, 2H), 1.48-1.18 (m, 6H), 1.16 (s, 6H).
*Partial presuppression of water signal using `watergate`
irradiation
[0193] .sup.13C NMR APT* (101 MHz, Methanol-d.sub.4) .delta.
179.21, 176.06, 174.25, 171.63, 165.07, 149.78, 148.77, 130.94,
126.88, 114.85, 111.80, 102.88, 96.10, 78.07, 76.09, 74.36, 71.28,
69.33, 59.00, 54.22, 53.31, 52.07, 44.77, 44.06, 41.76, 40.88,
40.54, 29.27, 29.22, 19.94, 19.10. *APT=Attached Proton Test
[0194] IR (Diamond ATR, solid) cm.sup.-1 3467, 3384, 2970, 2051,
1762, 1737, 1708, 1655, 1607, 1533, 1509, 1494, 1469, 1440, 1376,
1349, 1333, 1292, 1258, 1230, 1208, 1167, 1147, 1121, 1080, 1032,
985, 942, 926, 888, 865, 820, 771, 754, 717, 690, 675, 648, 616,
563, 542, 513, 476. See FIG. 1.3
[0195] IR (Diamond ATR, film) cm.sup.-1 3422, 2964, 1742, 1656,
1598, 1506, 1490, 1440, 1373, 1266, 1224, 1169, 1084, 1033, 929,
709, 567, 511. See FIG. 1.3
[0196] A. Single Crystal X-Ray Diffraction (See FIGS. 2.4.1 and
2.4.2)
[0197] From a suspension in its mother liquor, a suitable single
crystal of size 0.55.times.0.48.times.0.4 mm was finally selected
and implemented on the diffractometer.
TABLE-US-00008 Structural data Empirical formula C.sub.34H.sub.49N
O.sub.15 Extended formula C.sub.29H.sub.40N O.sub.9,
C.sub.4H.sub.5O.sub.5, CH.sub.4O Formula weight 711.74 Temperature
150(2)K Wavelength 0.71073 .ANG. Crystal system, space group
orthorhombic, P 2.sub.1 2.sub.1 2.sub.1 Unit cell dimensions: a =
11.3958(8) .ANG., .alpha. = 90.degree. b = 15.5163(16) .ANG.,
.beta. = 90.degree. c = 19.3680(16) .ANG., .gamma. = 90.degree.
Volume 3424.7(5) .ANG..sup.3 Z, Calculated density 4, 1.38 (g
cm.sup.-1) Absorption coefficient 0.108 mm.sup.-1 F(000) 1520
Crystal size 0.55 .times. 0.48 .times. 0.4 mm Crystal color
colourless Theta range for data collection 3.06 to 27.47.degree.
h_min, h_max -14, 14 k_min, k_max -20, 11 l_min, l_max -25, 14
Reflections collected/unique 14680/4317 [.sup.aR(int) = 0.0515]
Reflections [I > 2.sigma.] 3608 Completeness to theta_max 0.989
Absorption correction type multi-scan Max. and min. transmission
0.958, 0.839 Refinement method: Full-matrix least-squares on
F.sup.2 Data/restraints/parameters 4317/0/474 .sup.bGoodness-of-fit
1.034 Final R indices [I > 2.sigma.]: .sup.cR.sub.1 = 0.0397,
.sup.dwR.sub.2 = 0.0825 R indices (all data): .sup.cR.sub.1 =
0.0531, .sup.dwR.sub.2 = 0.0878 Largest diff. peak and hole 0.26
and -0.238 e..ANG..sup.-3
Atomic coordinates, site occupancy (%) and equivalent isotropic
displacement parameters (A.sup.2.times.10.sup.3). U(eq) is defined
as one third of the trace of the orthogonalized Uij tensor.
[0198] Atom numbering of FIG. 2.4.1 corresponds to below table.
TABLE-US-00009 Atom x y z occ. U(eq) C1 0.8742(2) 0.06078(16)
0.19603(12) 1 0.0184(5) H1 0.8565 0.0356 0.2395 1 0.022 C2
0.8762(2) 0.01795(16) 0.13644(13) 1 0.0188(5) C3 0.9050(2)
0.07250(16) 0.07486(11) 1 0.0170(5) H3 0.9806 0.0529 0.0538 1 0.02
C4 0.9198(2) 0.16514(15) 0.10526(11) 1 0.0162(5) H4 1.0039 0.1809
0.098 1 0.019 C5 0.9039(2) 0.15418(16) 0.18536(12) 1 0.0174(5) C6
1.0098(2) 0.18290(17) 0.22700(12) 1 0.0235(6) H6A 1.037 0.2405
0.2119 1 0.028 H6B 1.0752 0.1413 0.2222 1 0.028 C7 0.9654(3)
0.18574(18) 0.30140(13) 1 0.0267(6) H7A 1.0095 0.2288 0.3288 1
0.032 H7B 0.9732 0.1286 0.3237 1 0.032 C8 0.8361(3) 0.21154(17)
0.29473(12) 1 0.0238(6) H8A 0.7851 0.1693 0.3186 1 0.029 H8B 0.8226
0.2692 0.3151 1 0.029 N9 0.81061(19) 0.21246(14) 0.21739(10) 1
0.0176(4) H9 0.825(3) 0.269(2) 0.2017(17) 1 0.05 C10 0.6851(2)
0.19196(17) 0.20232(13) 1 0.0218(6) H10A 0.635 0.2391 0.2201 1
0.026 H10B 0.6631 0.1385 0.227 1 0.026 C11 0.6617(2) 0.18021(15)
0.12509(12) 1 0.0191(5) H11A 0.6845 0.121 0.1116 1 0.023 H11B
0.5764 0.1864 0.1166 1 0.023 C12 0.7267(2) 0.24357(15) 0.07984(12)
1 0.0173(5) C13 0.8483(2) 0.23587(15) 0.07084(11) 1 0.0158(5) C14
0.9095(2) 0.29500(15) 0.02929(12) 1 0.0183(5) H14 0.9918 0.2903
0.0226 1 0.022 C15 0.8457(2) 0.35990(16) -0.00131(12) 1 0.0200(5)
C16 0.7266(2) 0.36742(16) 0.00808(13) 1 0.0208(5) C17 0.6642(2)
0.31099(15) 0.04811(12) 1 0.0188(5) H17 0.582 0.3171 0.0542 1 0.023
C18 0.7889(2) 0.4844(2) -0.04641(17) 1 0.0369(7) H18A 0.802 0.5311
-0.0125 1 0.044 H18B 0.7814 0.5104 -0.0929 1 0.044 C19 0.8404(3)
-0.11951(17) 0.18391(15) 1 0.0307(6) H19A 0.912 -0.1207 0.2119 1
0.046 H19B 0.8203 -0.1782 0.1695 1 0.046 H19C 0.7759 -0.0954 0.2113
1 0.046 C21 0.8370(2) 0.02556(14) -0.03526(12) 1 0.0158(5) C22
0.7342(2) 0.02954(15) -0.08511(12) 1 0.0162(5) C23 0.7016(2)
0.12332(15) -0.10139(12) 1 0.0187(5) H23A 0.6336 0.1238 -0.1332 1
0.022 H23B 0.6773 0.1523 -0.0582 1 0.022 C24 0.8000(2) 0.17318(15)
-0.13336(12) 1 0.0184(5) C25 0.8506(3) 0.2801(2) -0.21480(17) 1
0.0398(8) H25A 0.8814 0.321 -0.1808 1 0.06 H25B 0.8156 0.3119
-0.2534 1 0.06 H25C 0.9146 0.2438 -0.232 1 0.06 C31 0.6261(2)
-0.01467(15) -0.05365(12) 1 0.0180(5) H31A 0.599 0.02 -0.0139 1
0.022 H31B 0.5625 -0.0146 -0.0885 1 0.022 C32 0.6456(2)
-0.10722(15) -0.02934(12) 1 0.0181(5) H32A 0.7012 -0.1076 0.0099 1
0.022 H32B 0.6803 -0.1416 -0.0673 1 0.022 C33 0.5293(2)
-0.14767(15) -0.00701(13) 1 0.0196(5) H33A 0.4777 -0.151 -0.048 1
0.024 H33B 0.4913 -0.1082 0.0264 1 0.024 C34 0.5349(2) -0.23726(16)
0.02570(12) 1 0.0202(5) C35 0.4125(2) -0.26450(18) 0.04804(15) 1
0.0280(6) H35A 0.4172 -0.3188 0.0737 1 0.042 H35B 0.363 -0.2724
0.0072 1 0.042 H35C 0.3785 -0.2198 0.0777 1 0.042 C36 0.5878(3)
-0.30546(16) -0.02198(14) 1 0.0261(6) H36A 0.6674 -0.2881 -0.0353 1
0.039 H36B 0.5391 -0.3111 -0.0634 1 0.039 H36C 0.591 -0.3609 0.0022
1 0.039 O1 0.85935(16) -0.06660(11) 0.12358(9) 1 0.0246(4) O2
0.88492(17) 0.42476(12) -0.04504(10) 1 0.0291(5) HO2 0.832(3)
-0.035(2) -0.1469(17) 1 0.05 O3 0.68491(16) 0.43735(12)
-0.02943(10) 1 0.0313(5) O4 0.81179(14) 0.06809(10) 0.02407(8) 1
0.0175(4) O5 0.92726(15) -0.01197(11) -0.04651(9) 1 0.0229(4) O6
0.76585(17) -0.01235(11) -0.14790(8) 1 0.0213(4) O7 0.90141(17)
0.16881(12) -0.11593(10) 1 0.0289(4) O8 0.76172(16) 0.22615(12)
-0.18254(10) 1 0.0294(5) O9 0.60916(17) -0.22805(12) 0.08630(9) 1
0.0220(4) H09 0.624(3) -0.279(2) 0.1042(17) 1 0.05 C51 0.8226(2)
0.44762(17) 0.18302(13) 1 0.0236(6) C52 0.7112(2) 0.45571(16)
0.22623(13) 1 0.0224(6) H52 0.721 0.5059 0.2581 1 0.027 C53
0.6023(2) 0.47238(16) 0.18092(13) 1 0.0224(5) H53A 0.612 0.4418
0.1364 1 0.027 H53B 0.5323 0.4483 0.2042 1 0.027 C54 0.5820(2)
0.56740(17) 0.16673(12) 1 0.0207(5) O51 0.84243(18) 0.51249(12)
0.14213(10) 1 0.0327(5) O52 0.88827(16) 0.38509(12) 0.18781(10) 1
0.0289(4) O53 0.69947(19) 0.38065(13) 0.26660(10) 1 0.0341(5) H53
0.628(3) 0.377(2) 0.2833(17) 1 0.05 O54 0.48847(17) 0.60192(12)
0.18067(9) 1 0.0274(4) O55 0.67121(17) 0.60868(11) 0.14119(9) 1
0.0263(4) H55 0.757(3) 0.559(2) 0.1382(16) 1 0.05 C61 0.4392(3)
0.46918(18) 0.34036(15) 1 0.0321(7) H61A 0.3848 0.4648 0.3794 1
0.048 H61B 0.3981 0.4932 0.3003 1 0.048 H61C 0.5048 0.5069 0.3529 1
0.048 O62 0.4827(2) 0.38596(14) 0.32369(11) 1 0.0418(6) H62
0.457(3) 0.347(2) 0.3498(17) 1 0.05
[0199] B. X-Ray Powder Diffraction
[0200] The sample was pure, there is no doubt that this is the
correct phase. However there is a gap of certain diffraction lines,
which would be associated with a variation of unit cell parameters.
There may be a change in the rate of hydration for example, to
cause such a phenomenon (for view of diagrams and experimental
details, see FIG. 2.4.3).
Example 5: Preparation and Analyses of Homoharringtonine Hydrogen
(2S,3S)-Tartrate
##STR00011##
[0202] This ionic compound was obtained from commercial
homoharringtonine mixed with commercial (2S,3S-(-)-tartaric acid
(unnatural form) according to the general procedure, then isolated
as a white prismatic solid mp 202-205.degree. C. (uncorrected) from
MeOH. (198.1-203.9, measured by DSC, see FIG. 3.4). Several
potentially acceptable crystals were kept suspended in their mother
liquors for the subsequent X-ray diffraction analysis. (see
below).
[0203] DSC Analysis (See FIG. 3.4)
[0204] .sup.1H NMR (400 MHz, Methanol-d.sub.4)*.delta. 6.81 (s,
1H), 6.75 (s, 1H), 6.10 (d, J=9.5 Hz, 1H), 5.97 (d, J=1.1 Hz, 1H),
5.94 (d, J=1.1 Hz, 1H), 5.34 (s, 1H), 4.36 (s, 2H), 4.18 (d, J=9.6
Hz, 1H), 3.82 (s, 3H), 3.55 (s, 3H), 2.24 (d, J=16.2 Hz, 2H), 1.95
(d, J=16.1 Hz, 1H), 1.16 (s, 6H). *Partial presuppression of water
signal using `watergate` irradiation
[0205] .sup.13C NMR APT* (101 MHz, Methanol-d4) .delta. 176.81,
174.28, 171.67, 165.24, 149.87, 148.85, 130.83, 126.76, 114.91,
111.89, 102.93, 96.04, 78.34, 76.13, 74.38, 74.10, 71.32, 59.10,
54.28, 53.22, 52.12, 44.80, 44.09, 40.91, 40.46, 29.20, 29.19,
19.95, 19.13. *APT=Attached Proton Test
[0206] IR (Diamond ATR, solid) cm.sup.-1 3502, 3048, 2971, 2884,
2051, 1981, 1765, 1736, 1656, 1592, 1506, 1490, 1432, 1375, 1348,
1321, 1295, 1265, 1227, 1205, 1165, 1147, 1111, 1081, 1031, 984,
939, 921, 887, 866, 831, 810, 727, 691, 675, 615, 564, 510, 477.
See FIG. 1.4
[0207] IR (Diamond ATR, film) cm.sup.-1 3419, 2963, 1741, 1656,
1611, 1506, 1489, 1440, 1373, 1265, 1224, 1168, 1118, 1083, 1035,
983, 928, 674, 614, 512, 477. See FIG. 1.4
[0208] X-Ray Crystallographic Studies
[0209] A. Single Crystal X-Ray Diffraction (See FIGS. 2.5.1 and
2.5.2)
[0210] From a suspension in its mother liquor, a suitable single
crystal of size 0.35.times.0.28.times.0.19 mm was finally selected
and implemented on the diffractometer.
TABLE-US-00010 Structural data Empirical formula .sub.33H.sub.45N
O.sub.15 Extended formula C.sub.33H.sub.45N.sub.1 O.sub.15 Formula
weight 695.7 Temperature 150(2)K Wavelength 0.71073 .ANG. Crystal
system, space group orthorhombic, P 2.sub.1 2.sub.1 2.sub.1 Unit
cell dimensions a = 10.7962(3) .ANG., .alpha. = 90.degree. b =
16.3649(5) .ANG., .beta. = 90.degree. c = 18.6773(5) .ANG., .gamma.
= 90.degree. Volume 3299.88(16) .ANG..sup.3 Z, Calculated density
4, 1.4 (g cm.sup.-1) Absorption coefficient 0.111 mm.sup.-1 F(000)
1480 Crystal size 0.35 .times. 0.28 .times. 0.19 mm Crystal color
colourless Theta range for data collection 3.12 to 27.48.degree.
h_min, h_max -14, 14 k_min, k_max -18, 21 l_min, l_max -23, 24
Reflections collected/unique 53493/4200 [.sup.aR(int) = 0.0357]
Reflections [I > 2.sigma.] 4022 Completeness to theta_max 0.997
Absorption correction type multi-scan Max. and min. transmission
0.979, 0.921 Refinement method Full-matrix least-squares on F.sup.2
Data/restraints/parameters 4200/0/464 .sup.bGoodness-of-fit 1.044
Final R indices [I > 2.sigma.]: .sup.cR.sub.1 = 0.0289,
.sup.dwR.sub.2 = 0.0766 R indices (all data): .sup.cR.sub.1 =
0.0308, .sup.dwR.sub.2 = 0.0781 Largest diff, peak and hole: 0.245
and -0.156 e..ANG..sup.-3
Atomic coordinates, site occupancy (%) and equivalent isotropic
displacement parameters (A.sup.2.times.10.sup.3). U(eq) is defined
as one third of the trace of the orthogonalized Uij tensor. Atom
numbering of FIG. 2.5.1 corresponds to below table.
TABLE-US-00011 Atom x y z occ. U(eq) C1 0.60365(16) 0.56010(10)
0.77843(9) 1 0.0207(3) H1 0.6286 0.5375 0.7338 1 0.025 C2
0.59199(16) 0.51759(10) 0.83901(9) 1 0.0204(3) C3 0.54863(14)
0.56685(9) 0.90214(8) 1 0.0173(3) H3 0.4655 0.5475 0.9185 1 0.021
C4 0.53958(14) 0.65545(9) 0.87170(8) 1 0.0159(3) H4 0.4499 0.6705
0.8739 1 0.019 C5 0.57198(15) 0.64844(10) 0.79030(9) 1 0.0176(3) C6
0.46897(15) 0.68171(10) 0.74191(8) 1 0.0209(3) H6A 0.4358 0.7336
0.7612 1 0.025 H6B 0.4003 0.6418 0.738 1 0.025 C7 0.52972(18)
0.69568(12) 0.66864(9) 1 0.0279(4) H7A 0.4941 0.7444 0.6449 1 0.034
H7B 0.5177 0.6476 0.6372 1 0.034 C8 0.66784(17) 0.70863(10)
0.68489(9) 1 0.0232(3) H8A 0.7186 0.6657 0.6617 1 0.028 H8B 0.6957
0.7627 0.6674 1 0.028 N9 0.67903(13) 0.70357(8) 0.76564(7) 1
0.0175(3) H9 0.667(3) 0.7517(18) 0.7806(16) 1 0.05 C10 0.80715(16)
0.68107(11) 0.78842(9) 1 0.0233(3) H10A 0.8653 0.7246 0.7736 1
0.028 H10B 0.8321 0.6299 0.7641 1 0.028 C11 0.81574(15) 0.66918(10)
0.86982(9) 1 0.0211(3) H11A 0.7888 0.613 0.8818 1 0.025 H11B 0.9033
0.6749 0.8847 1 0.025 C12 0.73790(14) 0.72935(10) 0.91171(9) 1
0.0183(3) C13 0.60770(14) 0.72158(9) 0.91235(8) 1 0.0160(3) C14
0.53534(15) 0.77926(9) 0.94934(8) 1 0.0177(3) H14 0.4476 0.7753
0.9498 1 0.021 C15 0.59463(16) 0.84150(10) 0.98478(9) 1 0.0203(3)
C16 0.72235(16) 0.84894(10) 0.98391(9) 1 0.0228(3) C17 0.79564(15)
0.79481(11) 0.94736(9) 1 0.0215(3) H17 0.8831 0.8013 0.9461 1 0.026
C18 0.64308(19) 0.95977(12) 1.03596(11) 1 0.0325(4) H18A 0.6364
1.0053 1.0012 1 0.039 H18B 0.6407 0.9827 1.085 1 0.039 C19
0.6668(2) 0.39314(10) 0.79407(10) 1 0.0279(4) H19A 0.7405 0.4221
0.7771 1 0.042 H19B 0.6068 0.3884 0.7549 1 0.042 H19C 0.6903 0.3384
0.8106 1 0.042 C21 0.60705(15) 0.51652(9) 1.01819(8) 1 0.0167(3)
C22 0.72136(14) 0.50930(9) 1.06633(8) 1 0.0166(3) C23 0.77327(15)
0.59411(9) 1.08420(9) 1 0.0192(3) H23A 0.8471 0.5876 1.1151 1 0.023
H23B 0.8004 0.6208 1.0393 1 0.023 C24 0.68149(16) 0.64899(10)
1.12155(9) 1 0.0217(3) C25 0.6586(2) 0.76969(15) 1.18925(17) 1
0.0533(7) H25A 0.6067 0.7412 1.2243 1 0.08 H25B 0.6057 0.7964
1.1536 1 0.08 H25C 0.709 0.8109 1.2137 1 0.08 C31 0.82188(15)
0.46027(9) 1.02660(9) 1 0.0191(3) H31A 0.8479 0.4917 0.9839 1 0.023
H31B 0.8949 0.4551 1.0583 1 0.023 C32 0.78268(16) 0.37451(9)
1.00245(9) 1 0.0197(3) H32A 0.7604 0.3411 1.0447 1 0.024 H32B 0.709
0.3784 0.9711 1 0.024 C33 0.88905(17) 0.33394(10) 0.96190(10) 1
0.0233(3) H33A 0.9647 0.3386 0.9916 1 0.028 H33B 0.9038 0.3655
0.9175 1 0.028 C34 0.87253(17) 0.24391(10) 0.94135(9) 1 0.0230(3)
C35 0.98538(19) 0.21661(13) 0.89846(13) 1 0.0370(5) H35A 0.9902
0.2482 0.854 1 0.055 H35B 0.9779 0.1584 0.8871 1 0.055 H35C 1.0605
0.2258 0.9268 1 0.055 C36 0.8548(2) 0.18850(12) 1.00617(11) 1
0.0385(5) H36A 0.8438 0.132 0.9901 1 0.058 H36B 0.7814 0.2059
1.0329 1 0.058 H36C 0.9279 0.192 1.0372 1 0.058 O1 0.61196(13)
0.43788(7) 0.85237(7) 1 0.0255(3) O2 0.54324(13) 0.90276(8)
1.02592(7) 1 0.0284(3) O3 0.75632(13) 0.91539(8) 1.02482(7) 1
0.0320(3) O4 0.63780(10) 0.56087(7) 0.95949(6) 1 0.0183(2) O5
0.50856(11) 0.48612(7) 1.02953(7) 1 0.0244(3) O6 0.69079(12)
0.46715(7) 1.13040(6) 1 0.0221(2) HO6 0.616(3) 0.4684(17)
1.1363(15) 1 0.05 O7 0.57142(13) 0.64049(9) 1.12151(10) 1 0.0408(4)
O8 0.73882(13) 0.71156(8) 1.15402(8) 1 0.0352(3) O9 0.76355(13)
0.23916(8) 0.89708(8) 1 0.0302(3) HO9 0.751(3) 0.1897(18)
0.8886(15) 1 0.05 C51 0.69752(17) 1.00450(11) 0.84160(9) 1
0.0240(3) C52 0.75591(16) 0.92992(10) 0.80382(9) 1 0.0216(3) H52
0.8202 0.9054 0.8358 1 0.026 C53 0.81501(17) 0.95062(11) 0.73116(9)
1 0.0248(3) H53 0.7586 0.9885 0.7048 1 0.03 C54 0.94378(18)
0.99098(12) 0.73715(10) 1 0.0291(4) O51 0.76283(13) 1.06995(8)
0.84564(8) 1 0.0331(3) O52 0.59210(13) 0.99642(8) 0.86509(8) 1
0.0337(3) O53 0.65877(13) 0.87221(7) 0.79311(7) 1 0.0276(3) HO53
0.604(3) 0.8924(17) 0.8176(15) 1 0.05 O54 0.82858(17) 0.87829(10)
0.69067(9) 1 0.0424(4) HO54 0.905(3) 0.8812(18) 0.6760(15) 1 0.05
O55 1.02589(15) 0.96663(11) 0.69804(9) 1 0.0465(4) O56 0.95769(14)
1.05015(9) 0.78265(9) 1 0.0373(3) HO56 0.868(3) 1.0629(17)
0.8111(14) 1 0.05
[0211] B. X-Ray Powder Diffraction
[0212] The sample was pure. There was a very good match between the
experimental pattern and the calculated pattern (for view of
diagrams and experimental details, see FIG. 2.5.3).
Example 6: Preparation and Analyses of Homoharringtonine Hydrogen
(2R,3R)-Tartrate (Diastemomer of Example 5)
##STR00012##
[0214] This ionic compound was obtained from commercial
homoharringtonine mixed with commercial (+)-(2R,3R)-tartaric acid
according to the general procedure in which the solvent was
methanol, then isolated as a white prismatic solid mp
206-208.degree. C. (uncorrected) from MeOH. (204.6-208.5, measured
by DSC, see FIG. 3.5). Several potentially acceptable crystals were
kept suspended in their mother liquors for the subsequent X-ray
diffraction analysis. (see below).
[0215] DSC Analysis (See FIG. 3.5)
[0216] .sup.1H NMR (400 MHz, Methanol-d.sub.4)*.delta. 6.80 (s,
1H), 6.75 (s, 1H), 6.10 (d, J=9.7 Hz, 1H), 5.97 (d, J=1.0 Hz, 1H),
5.94 (d, J=1.1 Hz, 1H), 5.34 (s, 1H), 4.36 (s, 2H), 4.18 (d, J=9.6
Hz, 1H), 3.82 (s, 3H), 3.55 (s, 3H), 3.44-3.32 (m, 2H), 3.28-3.16
(m, 1H), 2.69 (dd, J=13.7, 5.9 Hz, 1H), 2.27-2.21 (m, 2H),
2.21-1.97 (m, 2H), 1.95 (d, J=16.1 Hz, 1H), 1.49-1.18 (m, 6H), 1.16
(s, 6H). *Partial presuppression of water signal using `watergate`
irradiation
[0217] .sup.13C NMR APT* (101 MHz, Methanol-d.sub.4) .delta.
176.80, 174.24, 171.62, 165.20, 149.84, 148.83, 130.81, 126.73,
114.86, 111.85, 102.89, 96.00, 78.29, 76.09, 74.34, 74.07, 71.28,
59.05, 54.22, 53.18, 52.07, 44.76, 44.05, 40.87, 40.43, 29.23,
29.17, 29.15, 19.91, 19.10. *APT=Attached Proton Test
[0218] IR (Diamond ATR, solid) cm.sup.-1 3491, 3044, 2969, 1762,
1737, 1654, 1587, 1506, 1489, 1464, 1431, 1375, 1320, 1295, 1259,
1229, 1210, 1172, 1149, 1107, 1082, 1028, 984, 940, 924, 866, 819,
804, 735, 690, 616, 565, 512, 476. See FIG. 1.5
[0219] IR (Diamond ATR, film) cm.sup.-1 3417, 2963, 1741, 1655,
1611, 1505, 1489, 1440, 1373, 1265, 1223, 1167, 1118, 1082, 1034,
983, 928, 769, 675, 614, 565, 510, 478, 0, 1031, 984, 939, 921,
887, 866, 831, 810, 727, 691, 675, 615, 564, 510. See FIG. 1.5
[0220] X-Ray Crystallographic Studies
[0221] A. Single Crystal X-Ray Diffraction (See FIGS. 2.6.1 and
2.6.2)
[0222] From a suspension in its mother liquor, a suitable single
crystal of size 0.54.times.0.41.times.0.34 mm was finally selected
and implemented on the diffractometer.
TABLE-US-00012 Structural data Empirical formula
C.sub.33H.sub.45NO.sub.15 Extended formula
C.sub.29H.sub.40NO.sub.9, C.sub.4H.sub.5O.sub.6 Formula weight
695.7 Temperature 150(2) K Wavelength 0.71073 .ANG. Crystal system,
space group orthorhombic, P 2.sub.1 2.sub.1 2.sub.1 Unit cell
dimensions: a = 10.6770(3) .ANG., .alpha. = 90.degree. b =
16.6169(6) .ANG., .beta. = 90.degree. c = 18.7442(7) .ANG., .gamma.
= 90.degree. Volume 3325.6(2) .ANG..sup.3 Z, Calculated density 4,
1.39 (g cm.sup.-1) Absorption coefficient 0.110 mm.sup.-1 F(000)
1480 Crystal size 0.54 .times. 0.41 .times. 0.34 mm Crystal color
colourless Theta range for data collection 3.11 to 27.48.degree.
h_min, h_max -13, 10 k_min, k_max -21, 19 l_min, l_max -23, 15
Reflections collected/unique 15282/4165 [.sup.aR(int) = 0.0328]
Reflections [I > 2.sigma.] 3523 Completeness to theta_max 0.979
Absorption correction type multi-scan Max. and min. transmission
0.963, 0.891 Refinement method Full-matrix least-squares on F.sup.2
Data/restraints/parameters 4165/0/458 .sup.bGoodness-of-fit 1.021
Final R indices [I > 2.sigma.]: .sup.cR.sub.1 = 0.0368,
.sup.dwR.sub.2 = 0.0759 R indices (all data): .sup.cR.sub.1 =
0.0498, .sup.dwR.sub.2 = 0.0816 Largest diff. peak and hole 0.23
and -0.195 e .ANG..sup.-3
Atomic coordinates, site occupancy (%) and equivalent isotropic
displacement parameters (A.sup.2.times.10.sup.3). U(eq) is defined
as one third of the trace of the orthogonalized Uij tensor.
[0223] Atom numbering of FIG. 2.6.1 corresponds to below table.
TABLE-US-00013 Atom x y z occ. U(eq) C1 0.8908(2) 0.06521(13)
0.73105(13) 1 0.0187(5) H1 0.8694 0.0434 0.7763 1 0.022 C2
0.9037(2) 0.02207(13) 0.67131(13) 1 0.0194(5) C3 0.9421(2)
0.06998(13) 0.60762(12) 1 0.0170(5) H3 1.0267 0.0525 0.5907 1 0.02
C4 0.94749(19) 0.15799(13) 0.63555(12) 1 0.0149(5) H4 1.0376 0.1742
0.6331 1 0.018 C5 0.91467(19) 0.15286(12) 0.71688(12) 1 0.0164(5)
C6 1.0130(2) 0.19306(14) 0.76376(13) 1 0.0210(5) H6A 1.0399 0.2449
0.7427 1 0.025 H6B 1.0873 0.1579 0.7691 1 0.025 C7 0.9504(2)
0.20687(17) 0.83590(14) 1 0.0310(6) H7A 0.9805 0.2574 0.858 1 0.037
H7B 0.9677 0.1616 0.8688 1 0.037 C8 0.8100(2) 0.21218(14)
0.81897(12) 1 0.0218(5) H8A 0.7635 0.1686 0.8434 1 0.026 H8B 0.7756
0.2646 0.8345 1 0.026 N9 0.80013(17) 0.20331(11) 0.73928(10) 1
0.0167(4) H9 0.808(3) 0.2532(19) 0.7212(17) 1 0.05 C10 0.6742(2)
0.17363(14) 0.71581(13) 1 0.0212(5) H10A 0.609 0.2121 0.7316 1
0.025 H10B 0.6569 0.1211 0.7386 1 0.025 C11 0.6677(2) 0.16426(14)
0.63473(13) 1 0.0200(5) H11A 0.6969 0.1096 0.622 1 0.024 H11B
0.5792 0.1689 0.6196 1 0.024 C12 0.7446(2) 0.22516(13) 0.59340(13)
1 0.0178(5) C13 0.87646(19) 0.22060(12) 0.59378(12) 1 0.0142(5) C14
0.9478(2) 0.27752(13) 0.55633(12) 1 0.0176(5) H14 1.0367 0.2753
0.5567 1 0.021 C15 0.8861(2) 0.33632(13) 0.51922(13) 1 0.0188(5)
C16 0.7568(2) 0.34102(14) 0.51918(13) 1 0.0229(5) C17 0.6842(2)
0.28758(13) 0.55610(13) 1 0.0210(5) H17 0.5955 0.2924 0.5566 1
0.025 C18 0.8325(3) 0.44907(16) 0.46298(16) 1 0.0358(7) H18A 0.8347
0.4683 0.413 1 0.043 H18B 0.8367 0.4963 0.4951 1 0.043 C19
0.8337(2) -0.10152(13) 0.71742(14) 1 0.0278(6) H19A 0.8918 -0.1024
0.7579 1 0.042 H19B 0.8166 -0.1568 0.702 1 0.042 H19C 0.7552
-0.0756 0.7319 1 0.042 C21 0.8802(2) 0.01342(12) 0.49504(13) 1
0.0172(5) C22 0.7657(2) 0.00350(13) 0.44714(12) 1 0.0171(5) C23
0.7075(2) 0.08447(12) 0.42747(13) 1 0.0185(5) H23A 0.637 0.0752
0.3942 1 0.022 H23B 0.6731 0.1097 0.4711 1 0.022 C24 0.7990(2)
0.14152(13) 0.39345(13) 1 0.0203(5) C25 0.8218(2) 0.26011(16)
0.32546(17) 1 0.0354(7) H25A 0.8661 0.2903 0.3626 1 0.053 H25B
0.7713 0.2972 0.2968 1 0.053 H25C 0.8828 0.2329 0.2947 1 0.053 C32
0.6115(2) -0.17869(13) 0.54608(14) 1 0.0218(5) H32A 0.5368 -0.179
0.5147 1 0.026 H32B 0.5885 -0.1491 0.59 1 0.026 C37 0.7146(2)
-0.13192(12) 0.50853(13) 1 0.0197(5) H37A 0.7423 -0.1619 0.4657 1
0.024 H37B 0.7874 -0.1262 0.5409 1 0.024 C38 0.6679(2) -0.04831(12)
0.48638(13) 1 0.0192(5) H38A 0.6402 -0.0191 0.5296 1 0.023 H38B
0.594 -0.0549 0.455 1 0.023 C42 0.6411(2) -0.26590(13) 0.56665(13)
1 0.0214(5) C44 0.5296(2) -0.30128(16) 0.60626(15) 1 0.0310(6) H44A
0.5486 -0.3567 0.6205 1 0.047 H44B 0.456 -0.301 0.575 1 0.047 H44C
0.5122 -0.2689 0.6488 1 0.047 C45 0.6740(3) -0.31800(14)
0.50279(15) 1 0.0351(7) H45A 0.7474 -0.2955 0.4785 1 0.053 H45B
0.603 -0.3193 0.4697 1 0.053 H45C 0.6928 -0.3728 0.5189 1 0.053 O1
0.88885(17) -0.05711(9) 0.65947(9) 1 0.0254(4) O2 0.93569(16)
0.39596(10) 0.47637(9) 1 0.0273(4) O3 0.71970(17) 0.40404(11)
0.47595(10) 1 0.0334(4) O4 0.85067(13) 0.06035(9) 0.55137(8) 1
0.0176(3) O5 0.97892(14) -0.01899(9) 0.48520(9) 1 0.0250(4) O6
0.80450(16) -0.03582(10) 0.38304(9) 1 0.0231(4) HO6 0.878(3)
-0.0464(19) 0.3873(17) 1 0.05 O7 0.91117(15) 0.13633(11)
0.39748(11) 1 0.0374(5) O8 0.74109(15) 0.20093(9) 0.35831(10) 1
0.0275(4) O9 0.74716(17) -0.26262(10) 0.61420(11) 1 0.0320(5) HO9
0.755(3) -0.307(2) 0.6285(18) 1 0.05 C51 0.5651(2) 0.49263(15)
0.76865(15) 1 0.0299(6) C52 0.6393(2) 0.42749(13) 0.72898(15) 1
0.0237(5) H52 0.6068 0.4235 0.6791 1 0.028 C53 0.7815(2)
0.44249(13) 0.72627(14) 1 0.022 H53 0.8101 0.4609 0.7744 1 0.026
C54 0.8215(2) 0.50558(14) 0.67017(14) 1 0.0241(5) O51 0.5872(2)
0.56776(10) 0.75398(12) 1 0.0416(5) HO51 0.663(3) 0.5688(18)
0.7178(18) 1 0.05 O52 0.48857(19) 0.47183(12) 0.81317(11) 1
0.0462(6) O53 0.61742(18) 0.35372(10) 0.76410(12) 1 0.0383(5) HO53
0.566(3) 0.361(2) 0.7939(18) 1 0.05 O54 0.84448(16) 0.36948(10)
0.70908(10) 1 0.0275(4) HO54 0.887(3) 0.3812(19) 0.6718(18) 1 0.05
O55 0.90625(17) 0.48767(11) 0.62938(10) 1 0.0342(4) O56 0.76291(16)
0.57314(9) 0.67107(11) 1 0.0323(4)
[0224] A. X-Ray Powder Diffraction
[0225] The sample was pure and there was a very good match between
the experimental pattern and the calculated pattern (for view of
diagrams and experimental details, see FIG. 2.6.3).
Example 7: Preparation and Analyses of Homoharringtonine Hydrogen
(2'''S)-Citramalate
##STR00013##
[0227] This ionic compound was obtained from commercial
homoharringtonine mixed with commercial (2S)-citramalic acid
according to the general procedure in which the solvent was
methanol, then isolated as a white prismatic solid mp
195.9-198.9.degree. C. (measured by DSC, see FIG. 3.6). Several
potentially acceptable crystals were kept suspended in their mother
liquors for the subsequent X-ray diffraction analysis. (see
below).
[0228] DSC Analysis (See FIG. 3.6)
[0229] .sup.1H NMR (400 MHz, Methanol-d.sub.4)*.delta. 6.80 (s,
1H), 6.74 (s, 1H), 6.09 (d, J=9.6 Hz, 1H), 5.96 (d, J=0.9 Hz, 1H),
5.93 (d, J=0.9 Hz, 1H), 5.33 (s, 1H), 4.17 (d, J=9.6 Hz, 1H), 3.81
(s, 3H), 3.54 (s, 3H), 3.45-3.31 (m, 2H), 3.19 (dd, J=10.6, 6.9 Hz,
1H), 2.70 (d, J=15.7 Hz, 2H), 2.63 (d, J=15.7 Hz, 1H), 2.26-2.12
(m, 4H), 1.94 (d, J=16.1 Hz, 2H), 1.45-1.29 (m, 9H), 1.29-1.17 (m,
1H), 1.15 (s, 6H). *Partial presuppression of water signal using
`watergate` irradiation
[0230] .sup.13C NMR (101 MHz, MeOD) .delta. 181.21, 176.08, 174.23,
171.62, 165.18, 149.81, 148.79, 130.84, 126.78, 114.87, 114.58,
111.53, 102.58, 95.71, 78.24, 76.08, 74.03, 73.18, 71.27, 58.75,
53.91, 52.90, 51.79, 48.63, 46.32, 44.47, 43.77, 40.59, 40.15,
28.94, 28.89, 28.87, 26.22, 19.62, 18.80.
[0231] IR (Diamond ATR, solid) cm.sup.-1 2965, 1759, 1739, 1710,
1651, 1506, 1489, 1371, 1341, 1225, 1162, 1079, 1033, 972, 944,
925, 885, 866, 830, 786, 714, 690, 643, 615, 584, 562, 511. See
FIG. 1.6
[0232] IR (Diamond ATR, film) cm.sup.-1 3434, 2968, 1744, 1656,
1590, 1505, 1490, 1374, 1265, 1224, 1166, 1084, 1033, 930, 710,
565. See FIG. 1.6
[0233] X-Ray Powder Diffraction
[0234] The powder sample is well crystallised, with a peak width of
0.102.degree. (2.theta.) at 17.597.degree. (2.theta.) (for view of
diagrams and experimental details, see FIG. 2.7.1).
Example 8: Preparation and Analyses of Homoharringtonine Hydrogen
(2'''R)-Citramalate
##STR00014##
[0236] This ionic compound was obtained from commercial
homoharringtonine mixed with commercial (2R)-citramalic acid
according to the general procedure in which the solvent was
methanol, then isolated as a white prismatic solid mp
202.7-204.7.degree. C. (measured by DSC, see FIG. 3.7). Several
potentially acceptable crystals were kept suspended in their mother
liquors for the subsequent X-ray diffraction analysis. (see
below).
[0237] DSC Analysis (See FIG. 3.7)
[0238] .sup.1H NMR (400 MHz, Methanol-d.sub.4)*.delta. 6.79 (s,
1H), 6.74 (s, 1H), 6.08 (d, J=9.6 Hz, 1H), 5.95 (d, J=1.0 Hz, 1H),
5.93 (d, J=1.0 Hz, 1H), 5.33 (s, 1H), 4.17 (d, J=9.6 Hz, 1H), 3.81
(s, 3H), 3.54 (s, 3H), 3.43-3.31 (m, 2H), 3.22-3.14 (m, 1H),
2.73-2.66 (m, 2H), 2.63 (d, J=15.7 Hz, 1H), 2.23 (d, J=16.0 Hz,
2H), 2.19 (s, 1H), 1.94 (d, J=16.1 Hz, 1H), 1.44-1.29 (m, 8H),
1.29-1.17 (m, 1H), 1.15 (s, 6H). *Partial presuppression of water
signal using `watergate` irradiation
[0239] .sup.13C NMR APT* (101 MHz, Methanol-d.sub.4) .delta.
181.21, 176.08, 174.23, 171.62, 165.18, 149.81, 148.79, 130.84,
126.78, 114.87, 111.82, 102.87, 96.00, 78.24, 76.08, 74.33, 73.18,
71.27, 59.04, 54.21, 53.20, 52.07, 48.94, 46.62, 44.76, 44.05,
40.87, 40.45, 29.23, 29.19, 29.17, 26.50, 19.92, 19.09.
*APT=Attached Proton Test
[0240] IR (Diamond ATR, solid) cm.sup.-1 3681, 3512, 2969, 2845,
1764, 1740, 1707, 1652, 1605, 1513, 1495, 1469, 1440, 1369, 1332,
1292, 1260, 1227, 1204, 1167, 1147, 1124, 1080, 1048, 1033, 1023,
991, 971, 930, 885, 869, 824, 786, 753, 718, 689, 676, 644, 614,
564, 512, 475. See FIG. 1.7
[0241] IR (Diamond ATR, film) cm.sup.-1 3434, 2968, 2845, 1742,
1655, 1582, 1506, 1490, 1458, 1374, 1265, 1224, 1166, 1084, 1047,
1033, 930, 831, 710, 565, 476. See FIG. 1.7
[0242] X-Ray Crystallographic Studies
[0243] A. Single Crystal X-Ray Diffraction (See FIGS. 2.8.1 and
2.8.2)
[0244] From a suspension in its mother liquor, a suitable single
crystal of size 0.44.times.0.32.times.0.16 mm was finally selected
and implemented on the diffractometer.
TABLE-US-00014 Structural data Empirical formula
C.sub.34H.sub.47NO.sub.14 Extended formula
C.sub.29H.sub.40NO.sub.9, C.sub.5H.sub.7O.sub.5 Formula weight
693.73 Temperature 150(2) K Wavelength 0.71073 .ANG. Crystal
system, space group orthorhombic, P 2.sub.1 2.sub.1 2.sub.1 Unit
cell dimensions a = 10.3550(3) .ANG., .alpha. = 90.degree. b =
17.0899(6) .ANG., .beta. = 90.degree. c = 19.2854(7) .ANG., .gamma.
= 90.degree. Volume 3412.9(2) .ANG..sup.3 Z, Calculated density 4,
1.35 (g cm.sup.-1) Absorption coefficient 0.105 mm.sup.-1 F(000)
1480 Crystal size 0.44 .times. 0.32 .times. 0.16 mm Crystal color
colourless Theta range for data collection 3.09 to 27.48.degree.
h_min, h_max -13, 13 k_min, k_max -22, 20 l_min, l_max -19, 25
Reflections collected/unique 16497/7790 [.sup.aR(int) = 0.0336]
Reflections [I > 2.sigma.] 6790 Completeness to theta_max 0.996
Absorption correction type multi-scan Max. and min. transmission
0.983, 0.880 Refinement method Full-matrix least-squares on F.sup.2
Data/restraints/parameters 7790/0/462 .sup.bGoodness-of-fit 1.026
Final R indices[I > 2.sigma.] .sup.cR.sub.1 = 0.0413,
.sup.dwR.sub.2 = 0.0899 R indices (all data) .sup.cR.sub.1 =
0.0508, .sup.dwR.sub.2 = 0.0948 Largest diff. peak and hole 0.403
and -0.199 e .ANG..sup.-3
Atomic coordinates, site occupancy (%) and equivalent isotropic
displacement parameters (A.sup.2.times.10.sup.3). U(eq) is defined
as one third of the trace of the orthogonalized Uij tensor.
[0245] Atom numbering of FIG. 2.8.1 corresponds to below table.
TABLE-US-00015 Atom x y z occ. U(eq) C1 0.11571(16) 0.05636(10)
0.26511(9) 1 0.0175(4) H1 0.1363 0.0342 0.2213 1 0.021 C2
0.09856(16) 0.01523(10) 0.32323(9) 1 0.0171(4) C3 0.06120(16)
0.06351(11) 0.38472(9) 1 0.0162(3) H3 -0.0279 0.0492 0.4002 1 0.019
C4 0.06272(15) 0.14871(10) 0.35705(9) 1 0.0143(3) H4 -0.0287 0.1675
0.3589 1 0.017 C5 0.09821(15) 0.14231(10) 0.27823(9) 1 0.0159(4) C6
0.00002(16) 0.18455(11) 0.23179(9) 1 0.0195(4) H6A -0.0761 0.1508
0.2234 1 0.023 H6B -0.0293 0.2336 0.254 1 0.023 C7 0.06914(19)
0.20226(14) 0.16369(10) 1 0.0282(5) H7A 0.0457 0.255 0.1465 1 0.034
H7B 0.046 0.1632 0.1279 1 0.034 C8 0.21369(17) 0.19809(11)
0.18060(9) 1 0.0190(4) H8A 0.2545 0.153 0.1569 1 0.023 H8B 0.2581
0.2467 0.166 1 0.023 N9 0.22010(13) 0.18836(9) 0.25825(7) 1
0.0156(3) HN9 0.215(2) 0.2413(15) 0.2762(14) 1 0.05 C10 0.34708(16)
0.15611(11) 0.28115(9) 1 0.0187(4) H10A 0.4167 0.193 0.2681 1 0.022
H10B 0.3634 0.1059 0.2571 1 0.022 C11 0.35010(16) 0.14274(11)
0.35966(9) 1 0.0182(4) H11A 0.313 0.0906 0.3698 1 0.022 H11B 0.4411
0.1423 0.3753 1 0.022 C12 0.27690(16) 0.20378(10) 0.40075(9) 1
0.0161(3) C13 0.14095(15) 0.20673(10) 0.39833(9) 1 0.0142(3) C14
0.07316(17) 0.26533(10) 0.43397(9) 1 0.0168(4) H14 -0.0183 0.2688
0.4315 1 0.02 C15 0.14387(18) 0.31747(10) 0.47263(9) 1 0.0200(4)
C16 0.27616(18) 0.31308(11) 0.47618(9) 1 0.0207(4) C17 0.34575(17)
0.25856(11) 0.43994(9) 1 0.0190(4) H17 0.4374 0.258 0.4414 1 0.023
C18 0.2137(2) 0.41952(12) 0.53342(11) 1 0.0334(5) H18A 0.2207
0.4679 0.5055 1 0.04 H18B 0.2103 0.4343 0.583 1 0.04 C19 0.1546(2)
-0.10852(11) 0.27911(11) 1 0.0300(5) H19A 0.0911 -0.1088 0.2414 1
0.045 H19B 0.1691 -0.1622 0.2952 1 0.045 H19C 0.2361 -0.0864 0.2622
1 0.045 C21 0.11247(16) 0.01321(10) 0.49827(9) 1 0.0161(3) C22
0.22813(16) 0.00398(10) 0.54736(9) 1 0.0178(4) C23 0.29449(17)
0.08219(10) 0.56296(10) 1 0.0189(4) H23A 0.369 0.0729 0.594 1 0.023
H23B 0.3277 0.1048 0.5192 1 0.023 C24 0.20436(17) 0.13941(10)
0.59644(10) 1 0.0196(4) C25 0.18690(19) 0.25170(12) 0.66695(12) 1
0.0293(5) H25A 0.1478 0.2853 0.6315 1 0.044 H25B 0.2392 0.2837
0.6984 1 0.044 H25C 0.1187 0.2253 0.6933 1 0.044 C31 0.32662(17)
-0.05075(11) 0.51274(10) 1 0.0204(4) H31A 0.361 -0.0244 0.4709 1
0.025 H31B 0.3997 -0.0586 0.5451 1 0.025 C32 0.27492(18)
-0.13115(11) 0.49143(10) 1 0.0222(4) H32A 0.2495 -0.1609 0.5333 1
0.027 H32B 0.1973 -0.1245 0.462 1 0.027 C33 0.37715(18)
-0.17662(11) 0.45161(11) 1 0.0248(4) H33A 0.4587 -0.1741 0.4783 1
0.03 H33B 0.3923 -0.1495 0.407 1 0.03 C34 0.34851(18) -0.26269(11)
0.43594(11) 1 0.0240(4) C36 0.4577(2) -0.29453(13) 0.39087(14) 1
0.0423(6) H36A 0.4411 -0.3497 0.3801 1 0.063 H36B 0.5397 -0.2899
0.4159 1 0.063 H36C 0.4623 -0.2644 0.3477 1 0.063 C35 0.3354(2)
-0.31160(13) 0.50108(12) 1 0.0363(5) H35A 0.2616 -0.2929 0.5283 1
0.054 H35B 0.4144 -0.3071 0.5288 1 0.054 H35C 0.3216 -0.3665 0.4884
1 0.054 O1 0.10699(13) -0.06191(7) 0.33525(7) 1 0.0244(3) O2
0.32223(14) 0.37011(8) 0.52078(7) 1 0.0304(3) O3 0.09930(13)
0.37738(8) 0.51464(7) 1 0.0288(3) O4 0.15171(11) 0.05193(7)
0.44075(6) 1 0.0171(3) O5 0.00628(12) -0.01257(8) 0.50746(7) 1
0.0222(3) O6 0.18733(13) -0.03191(8) 0.60999(7) 1 0.0221(3) HO6
0.120(3) -0.0138(16) 0.6222(14) 1 0.05 O7 0.08889(13) 0.13801(9)
0.59158(9) 1 0.0357(4) O8 0.26816(12) 0.19381(8) 0.63389(7) 1
0.0243(3) O9 0.22961(14) -0.26485(9) 0.39752(9) 1 0.0348(4) HO9
0.203(3) -0.3138(17) 0.3982(14) 1 0.05 C51 0.17383(18) 0.50529(11)
0.36479(10) 1 0.0247(4) C52 0.31671(18) 0.48833(12) 0.35581(12) 1
0.0299(5) H52A 0.3411 0.4469 0.3892 1 0.036 H52B 0.3654 0.5361
0.3686 1 0.036 C53 0.36065(18) 0.46262(12) 0.28366(12) 1 0.0288(5)
C54 0.2995(2) 0.51004(15) 0.22598(13) 1 0.0470(6) H54A 0.3131
0.5659 0.2348 1 0.071 H54B 0.2067 0.4991 0.2242 1 0.071 H54C 0.3392
0.4958 0.1816 1 0.071 C55 0.33695(18) 0.37452(11) 0.26995(10) 1
0.0243(4) O51 0.13621(14) 0.56592(9) 0.39014(9) 1 0.0361(4) O52
0.09277(13) 0.45011(9) 0.34618(8) 1 0.0300(3) HO52 0.147(2)
0.4107(15) 0.3218(14) 1 0.05 O53 0.49802(14) 0.47478(10)
0.28234(10) 1 0.0438(4) HO53 0.530(2) 0.4274(17) 0.2667(14) 1 0.05
O54 0.42729(14) 0.33586(9) 0.24647(8) 1 0.0368(4) O55 0.22436(12)
0.34821(8) 0.28413(7) 1 0.0261(3)
[0246] A. X-Ray Powder Diffraction
[0247] The sample was pure and well crystallised, with a peak width
of 0.107.degree. (2.theta.) at 16.992.degree. (2.theta.). There was
a very good match between the experimental pattern and the
calculated pattern (for view of diagrams and experimental details,
see FIG. 2.8.3).
Example 9: Preparation and Analyses of Homoharringtonine Hydrogen
Succinate
##STR00015##
[0249] This ionic compound was obtained from commercial
homoharringtonine mixed with commercial succinic acid according to
the general procedure in which the solvent was methanol, then
isolated as a white prismatic solid mp 158.1-160.0.degree. C.
(measured by DSC, see FIG. 3.8).
[0250] DSC Analysis (See FIG. 3.8)
[0251] .sup.1H NMR (400 MHz, Methanol-d.sub.4)*.delta. 6.77 (s,
1H), 6.71 (s, 1H), 6.07 (dd, J=9.6, 0.7 Hz, 1H), 5.95 (d, J=1.1 Hz,
1H), 5.92 (d, J=1.1 Hz, 1H), 5.31 (d, J=0.6 Hz, 1H), 4.12 (d, J=9.6
Hz, 1H), 3.79 (s, 3H), 3.54 (s, 3H), 2.49 (s, 4H), 2.22 (d, J=16.2
Hz, 1H), 1.93 (d, J=16.1 Hz, 2H), 1.15 (s, 6H). *Partial
presuppression of water signal using `watergate` irradiation
[0252] .sup.13 C NMR APT* (101 MHz, D.sub.2O) .delta. 179.49,
174.22, 171.93, 162.89, 147.84, 146.75, 129.74, 125.23, 113.38,
111.12, 101.62, 95.53, 76.99, 75.26, 73.68, 71.33, 58.41, 52.95,
52.23, 51.27, 48.86, 47.58, 42.72, 42.55, 39.19, 38.77, 31.20,
27.59, 18.59, 17.69. *APT=Attached Proton Test
[0253] IR (KBr, solid), cm.sup.-1 3571.1, 3375.3, 3083.4, 2964.4,
1755.4, 1736.7, 1661.8, 1575.5, 1504.8, 1489.7, 1375.0, 1346.3,
1326.1, 1267.2, 1227.1, 1188.3, 1151.7, 1083.4, 1034.7, 929.4,
859.4, 802.8, 758.1, 709.9, 658.9, 617.9, 561.2, 510.6. See FIG.
1.8
Example 10: Preparation and Analyses of
(3S,4S,5R,2'R)-Homoharringtonine Hydrogen Itaconate
##STR00016##
[0255] This ionic compound was obtained from commercial
homoharringtonine mixed with commercial itaconic acid according to
the general procedure in which the solvent was methanol, then
isolated as a white prismatic solid mp 178.3-181.2.degree. C.
(measured by DSC, see FIG. 3.9). Several potentially acceptable
crystals were kept suspended in their mother liquors for the
subsequent X-ray diffraction analysis. (see below).
[0256] DSC Analysis (See FIG. 3.9)
[0257] .sup.1H NMR (400 MHz, Methanol-d.sub.4)*.delta. 6.78 (s,
1H), 6.72 (s, 1H), 6.08 (d, J=9.6 Hz, 1H), 6.01 (d, J=1.7 Hz, 1H),
5.95 (d, J=1.1 Hz, 1H), 5.92 (d, J=1.1 Hz, 1H), 5.51 (q, J=1.2 Hz,
1H), 5.32 (s, 1H), 4.14 (d, J=9.6 Hz, 1H), 3.80 (s, 3H), 3.54 (s,
3H), 3.51-3.42 (m, 1H), 3.25-3.07 (m, 2H), 2.72-2.60 (m, 1H),
2.26-2.20 (m, 2H), 2.20-2.07 (m, 2H), 1.94 (d, J=16.1 Hz, 2H),
1.47-1.29 (m, 5H), 1.23 (d, J=10.6 Hz, 1H), 1.15 (s, 6H). *Partial
presuppression of water signal using `watergate` irradiation
[0258] .sup.13C NMR (101 MHz, MeOD)** .delta. 125.11, 114.53,
111.47, 102.52, 96.09, 74.12, 58.66, 53.95, 53.18, 48.70, 44.48,
43.78, 41.66, 40.59, 40.44, 29.12, 28.94, 28.87, 19.70, 18.81.
**DEPT135: Distortionless Enhancement by Polarization Transfer
(non-quaternary carbons only)
[0259] IR (Diamond ATR, solid) cm.sup.-1 3473.2, 2968.4, 2899.8,
2564.7, 1760.7, 1733.5, 1657.6, 1569.7, 1506.4, 1488.9, 1436.1,
1374.9, 1348.9, 1264.2, 1240.7, 1226.2, 1185, 1168.8, 1149.8,
1112.1, 1082.4, 1043.1, 1032.9, 1022.2, 982, 928.1, 890, 866.7,
819.8, 772.1, 722.1, 690.1, 616.7, 543. See FIG. 1.9
[0260] IR (ATR, film) cm.sup.-1 3458.8, 2967, 1741.5, 1654.8,
1576.7, 1505.2, 1489.3, 1464.3, 1373.4, 1223.8, 1167, 1083.1,
1033.3, 933.6, 563.4. See FIG. 1.9
[0261] X-Ray Crystallographic Studies
[0262] Single Crystal X-Ray Diffraction (See FIGS. 2.9.1 and
2.9.2)
[0263] From a suspension in its mother liquor, a suitable single
crystal of size 0.39.times.0.22.times.0.1 mm was finally selected
and implemented on the diffractometer.
TABLE-US-00016 Structural data Empirical formula
C.sub.34H.sub.45NO.sub.13 Extended formula
C.sub.29H.sub.40NO.sub.9, C.sub.5H.sub.5O.sub.4 Formula weight
675.71 Temperature 150(2) K Wavelength 0.71073 .ANG. Crystal
system, space group orthorhombic, P 2.sub.1 2.sub.1 2.sub.1 Unit
cell dimensions a = 10.9895(4) .ANG., .alpha. = 90.degree. b =
16.1963(6) .ANG., .beta. = 90.degree. c = 18.7277(5) .ANG., .gamma.
= 90.degree. Volume 3333.33(19) .ANG..sup.3 Z, Calculated density
4, 1.346 (g cm.sup.-1) Absorption coefficient 0.103 mm.sup.-1
F(000) 1440 Crystal size 0.39 .times. 0.22 .times. 0.1 mm Crystal
color colourless Theta range for data collection 3.12 to
27.48.degree. h_min, h_max -11, 14 k_min, k_max -13, 20 l_min,
l_max -16, 24 Reflections collected/unique 15962/4207 [.sup.aR(int)
= 0.0449] Reflections [I > 2.sigma.] 3444 Completeness to
theta_max 0.986 Absorption correction type multi-scan Max. and min.
transmission 0.990, 0.844 Refinement method: Full-matrix
least-squares on F.sup.2 Data/restraints/parameters 4207/0/446
.sup.bGoodness-of-fit 1.054 Final R indices [I > 2.sigma.]
.sup.cR.sub.1 = 0.0411, .sup.dwR.sub.2 = 0.0914 R indices (all
data) .sup.cR.sub.1 = 0.0557, .sup.dwR.sub.2 = 0.0986 Largest diff.
peak and hole 0.312 and -0.249 e .ANG..sup.-3
Atomic coordinates, site occupancy (%) and equivalent isotropic
displacement parameters (A.sup.2.times.10.sup.3). U(eq) is defined
as one third of the trace of the orthogonalized Uij tensor.
[0264] Atom numbering of FIG. 2.9.1 corresponds to below table.
TABLE-US-00017 Atom x y z occ. U(eq) C1 0.4085(2) 0.94954(16)
0.28454(13) 1 0.0246(6) H1 0.3897 0.972 0.239 1 0.029 C2 0.4040(2)
0.99163(16) 0.34531(13) 1 0.0239(5) C3 0.4399(2) 0.94237(16)
0.40941(12) 1 0.0206(5) H3 0.5169 0.9646 0.4305 1 0.025 C4
0.4605(2) 0.85392(16) 0.38019(11) 1 0.0187(5) H4 0.548 0.8406
0.3893 1 0.022 C5 0.4471(2) 0.86236(16) 0.29702(12) 1 0.0216(5) C6
0.5613(2) 0.83684(18) 0.25641(12) 1 0.0244(6) H6A 0.5945 0.7845
0.2755 1 0.029 H6B 0.6247 0.8801 0.2599 1 0.029 C7 0.5195(3)
0.8261(2) 0.17935(13) 1 0.0330(7) H7A 0.5696 0.7843 0.1544 1 0.04
H7B 0.5244 0.879 0.153 1 0.04 C8 0.3878(3) 0.7973(2) 0.18586(13) 1
0.0347(7) H8A 0.3332 0.8339 0.1582 1 0.042 H8B 0.3791 0.7402 0.1675
1 0.042 N9 0.35665(19) 0.80066(14) 0.26453(10) 1 0.0225(5) H9
0.3728 0.749 0.284 1 0.027 C10 0.2240(2) 0.81867(19) 0.27555(14) 1
0.0293(6) H10A 0.1752 0.7717 0.2576 1 0.035 H10B 0.2013 0.8683
0.2477 1 0.035 C11 0.1946(2) 0.83313(19) 0.35427(13) 1 0.0280(6)
H11A 0.2153 0.8908 0.3668 1 0.034 H11B 0.1061 0.8258 0.3617 1 0.034
C12 0.2625(2) 0.77534(17) 0.40351(12) 1 0.0218(5) C13 0.3876(2)
0.78580(16) 0.41518(12) 1 0.0197(5) C14 0.4503(2) 0.73128(16)
0.45992(12) 1 0.0226(5) H14 0.5349 0.7378 0.4685 1 0.027 C15
0.3868(2) 0.66846(18) 0.49096(13) 1 0.0281(6) C16 0.2644(2)
0.65757(18) 0.47829(14) 1 0.0300(6) C17 0.1998(2) 0.70918(17)
0.43475(13) 1 0.0274(6) H17 0.1156 0.7006 0.426 1 0.033 C18
0.3315(3) 0.5504(2) 0.5409(2) 1 0.0595(10) H18A 0.3503 0.5025
0.5101 1 0.071 H18B 0.3207 0.5305 0.5905 1 0.071 C19 0.3222(3)
1.11383(18) 0.29761(14) 1 0.0333(7) H19A 0.2562 1.0809 0.277 1 0.05
H19B 0.3856 1.1225 0.2616 1 0.05 H19C 0.2902 1.1674 0.3132 1 0.05
C21 0.3633(2) 0.98532(16) 0.52334(12) 1 0.0193(5) C22 0.2491(2)
0.98238(16) 0.56983(12) 1 0.0210(5) C23 0.2120(2) 0.89286(16)
0.58578(12) 1 0.0225(5) H23A 0.1406 0.8932 0.618 1 0.027 H23B
0.1873 0.8659 0.5406 1 0.027 C24 0.3118(2) 0.84293(17) 0.61968(12)
1 0.0228(5) C25 0.3602(3) 0.7333(2) 0.69781(18) 1 0.0448(8) H25A
0.4231 0.7681 0.7195 1 0.067 H25B 0.3973 0.6974 0.6618 1 0.067 H25C
0.3218 0.6994 0.7348 1 0.067 C31 0.1446(2) 1.02575(17) 0.53105(13)
1 0.0239(5) H31A 0.1214 0.9918 0.4892 1 0.029 H31B 0.0735 1.0275
0.5635 1 0.029 C32 0.1704(2) 1.11351(17) 0.50526(13) 1 0.0272(6)
H32A 0.1946 1.1483 0.5463 1 0.033 H32B 0.2387 1.1127 0.4708 1 0.033
C33 0.0585(3) 1.1503(2) 0.46982(16) 1 0.0367(7) H33A -0.0106 1.1443
0.5033 1 0.044 H33B 0.0395 1.1164 0.4272 1 0.044 C34 0.0638(3)
1.2404(2) 0.44632(14) 1 0.0348(7) C35 -0.0534(3) 1.2636(3)
0.4090(2) 1 0.0667(12) H35A -0.0642 1.2289 0.3667 1 0.1 H35B
-0.0499 1.3217 0.3946 1 0.1 H35C -0.122 1.2554 0.4417 1 0.1 C36
0.0885(3) 1.2991(2) 0.50777(16) 1 0.0518(9) H36A 0.0919 1.3558
0.4897 1 0.078 H36B 0.1664 1.2849 0.5301 1 0.078 H36C 0.0232 1.2944
0.5432 1 0.078 O1 0.37336(17) 1.07063(11) 0.35827(9) 1 0.0295(4) O2
0.2228(2) 0.58991(14) 0.51681(12) 1 0.0481(6) O3 0.4284(2)
0.60914(13) 0.53788(11) 1 0.0427(5) O4 0.34267(14) 0.94422(11)
0.46170(8) 1 0.0201(4) O5 0.45495(15) 1.02085(12) 0.53858(9) 1
0.0266(4) O6 0.27352(16) 1.02392(12) 0.63524(9) 1 0.0251(4) HO6
0.342(3) 1.033(2) 0.6369(17) 1 0.05 O7 0.41851(16) 0.85198(13)
0.60753(11) 1 0.0356(5) O8 0.26888(16) 0.78534(13) 0.66423(11) 1
0.0366(5) O9 0.1644(2) 1.24749(16) 0.39741(12) 1 0.0513(6) HO9
0.160(3) 1.307(2) 0.3826(17) 1 0.05 C51 0.1037(3) 0.4553(2)
0.31090(15) 1 0.0388(7) C52 0.1159(3) 0.5480(2) 0.3010(2) 1
0.0512(9) H52A 0.0462 0.5679 0.2723 1 0.061 H52B 0.1112 0.5749
0.3484 1 0.061 C53 0.2325(3) 0.57416(19) 0.26485(16) 1 0.0403(8)
C54 0.3464(3) 0.57891(18) 0.30905(13) 1 0.0285(6) C55 0.2404(5)
0.5906(2) 0.19500(19) 1 0.0747(14) H55A 0.3166 0.6049 0.1745 1 0.09
H55B 0.1697 0.5881 0.1659 1 0.09 O51 0.00971(18) 0.41958(16)
0.29455(13) 1 0.0529(6) O52 0.19701(19) 0.41745(13) 0.33812(11) 1
0.0377(5) HO52 0.262(3) 0.454(2) 0.3511(17) 1 0.05 O53 0.41850(17)
0.63675(12) 0.30044(10) 1 0.0344(5) O54 0.36137(17) 0.52002(13)
0.35372(10) 1 0.0346(5)
Example 11: Preparation and Analyses of Homoharringtonine Hydrogen
Fumarate
##STR00017##
[0266] This ionic compound was obtained from commercial
homoharringtonine mixed with commercial fumaric acid according to
the general procedure in which the solvent was methanol, then
isolated as a white prismatic solid mp 103.5-107.2.degree. C.
(measured by DSC, see FIG. 3.10).
[0267] DSC Analysis (See FIG. 3.10)
[0268] .sup.1H NMR (400 MHz, Methanol-d.sub.4)*.delta. 6.80 (s,
1H), 6.74 (s, 1H), 6.65 (s, 2H), 6.09 (d, J=9.6 Hz, 1H), 5.96 (d,
J=0.9 Hz, 1H), 5.93 (d, J=0.9 Hz, 1H), 5.33 (s, 1H), 4.17 (d, J=9.6
Hz, 1H), 3.82 (s, 3H), 3.55 (s, 3H), 3.43-3.32 (m, 2H), 3.24-3.10
(m, 1H), 2.75-2.61 (m, 1H), 2.30-2.08 (m, 4H), 1.95 (d, J=16.1 Hz,
2H), 1.47-1.30 (m, 5H), 1.16 (s, 6H). *Partial presuppression of
water signal using `watergate" irradiation
[0269] .sup.13C NMR (101 MHz, MeOD)** .delta. 135.91, 114.59,
111.56, 102.57, 95.74, 74.03, 58.74, 53.89, 52.92, 51.79, 49.56,
48.62, 44.47, 43.77, 40.59, 40.16, 28.94, 28.90, 28.88, 19.64,
18.81. **DEPT135: Distortionless Enhancement by Polarization
Transfer (non-quaternary carbons only)
[0270] IR (ATR, solid), cm.sup.-1 3607.9, 3212.6, 2955.6, 1980.4,
1777.4, 1731.4, 1708.1, 1653.6, 1584.3, 1505.9, 1488.6, 1440.0,
1372.4, 1338.6, 1292.0, 1251.1, 1221.1, 1173.3, 1150.9, 1119.3,
1088.7, 1034.0, 982.0, 934.1, 903.3, 839.6, 790.3, 761.8, 646.0,
613.5, 563.6, 510.2. See FIG. 1.10
[0271] X-Ray Powder Diffraction
[0272] The powder sample is well crystallised, with a peak width of
0.119.degree. (2.theta.) at 19.564.degree. (2.theta.) (for view of
diagrams and experimental details, see FIG. 2.7.1).
Example 12: Preparation and Analyses of Homoharringtonine Hydrogen
Tartronate
##STR00018##
[0274] This ionic compound was obtained from commercial
homoharringtonine mixed with commercial tartronic acid according to
the general procedure in which the solvent was methanol, then
isolated as a white prismatic solid mp 163.1-167.6.degree. C.
(measured by DSC, see FIG. 3.11).
[0275] DSC Analysis (See FIG. 3.11)
[0276] .sup.1H NMR (400 MHz, Methanol-d.sub.4)*.delta. 6.80 (s,
1H), 6.75 (s, 1H), 6.09 (d, J=9.6 Hz, 1H), 5.96 (d, J=0.9 Hz, 1H),
5.94 (d, J=0.9 Hz, 1H), 5.34 (s, 1H), 4.18 (d, J=9.6 Hz, 1H), 3.82
(s, 3H), 3.54 (s, 3H), 2.69 (m, 1H), 2.22 (m, 4H), 2.04-1.91 (m,
2H), 1.47-1.29 (m, 5H), 1.23 (m, 1H), 1.15 (s, 6H). *Partial
presuppression of water signal using `watergate" irradiation
[0277] .sup.13C NMR (101 MHz, MeOD)** .delta. 114.60, 111.55,
102.61, 95.67, 74.02, 58.77, 53.94, 52.86, 51.79, 48.67, 44.47,
43.77, 40.59, 40.10, 28.94, 28.88, 28.84, 19.63, 18.80. **DEPT135:
Distortionless Enhancement by Polarization Transfer (non-quaternary
carbons only)
[0278] IR (Diamond ATR, solid) cm.sup.-1 3451, 2969, 2898, 2051,
1763, 1730, 1657, 1507, 1490, 1467, 1437, 1376, 1352, 1316, 1294,
1266, 1228, 1208, 1186, 1148, 1126, 1083, 1032, 1002, 985, 943,
927, 891, 866, 802, 753, 720, 690, 675, 652, 614, 563, 510, 477.
See FIG. 1.11
[0279] IR (Diamond ATR, film) cm.sup.-1 3429, 2965, 1744, 1655,
1505, 1489, 1440, 1374, 1266, 1224, 1165, 1084, 1033, 928, 807,
615. See FIG. 1.11
Example 13: Preparation and Analyses of Homoharringtonine Hydrogen
Malonate
##STR00019##
[0281] This ionic compound was obtained from commercial
homoharringtone mixed with commercial (2R)-citramalic acid
according to the general procedure in which the solvent was
methanol-d.sub.4, then isolated as a white prismatic solid mp
127.0-131.9.degree. C. (measured by DSC, see FIG. 3.12).
[0282] DSC Analysis (See FIG. 3.12)
[0283] .sup.1H NMR (400 MHz, Methanol-d.sub.4)*.delta. 6.81 (s,
1H), 6.75 (s, 1H), 6.10 (d, J=9.6 Hz, 1H), 5.97 (d, J=1.1 Hz, 1H),
5.94 (d, J=1.0 Hz, 1H), 5.34 (s, 1H), 4.18 (d, J=9.6 Hz, 1H), 3.82
(s, 3H), 3.55 (s, 3H), 3.24-3.17 (m, 1H), 2.74-2.64 (m, 1H),
2.30-2.09 (m, 4H), 1.95 (d, J=16.1 Hz, 2H), 1.48-1.30 (m, 5H), 1.16
(s, 6H). *Partial presuppression of water signal using `watergate`
irradiation
[0284] .sup.13C NMR APT* (101 MHz, Methanol-d4) .delta. 174.83,
174.22, 171.62, 165.26, 149.82, 148.81, 130.79, 126.75, 114.88,
111.83, 102.90, 95.90, 78.32, 76.09, 74.30, 71.27, 59.04, 54.21,
53.18, 52.07, 48.94, 44.75, 44.05, 40.87, 40.42, 29.22, 29.17,
29.14, 19.91, 19.09. See FIG. 1.12 *APT=Attached Proton Test
[0285] IR (Diamond ATR, solid) cm.sup.-1 3453.1, 2967.5, 2933.2,
2899.3, 1765.0, 1735.2, 1654.6, 1505.9, 1489.1, 1463.7, 1439.1,
1374.9, 1349.7, 1292.0, 1266.2, 1226.5, 1207.5, 1148.4, 1083.3,
1060.6, 1032.3, 1002.1, 985.4, 944.1, 925.5, 891.0, 858.5, 830.6,
797.6, 756.7, 721.5, 710.8, 690.8, 615.1, 565.1, 510.8, 498.3,
489.9, 478.9, 472.8. See FIG. 1.12
Example 14: Preparation and Analyses of Homoharringtonine
Dihydrogen Citrate
##STR00020##
[0287] This ionic compound was obtained from commercial
homoharringtonine mixed with commercial citric acid according to
the general procedure in which the solvent was methanol, then
isolated as a white prismatic solid mp 170.35-173.9.degree. C.
(measured by DSC, see FIG. 3.13). Several potentially acceptable
crystals were kept suspended in their mother liquors for the
subsequent X-ray diffraction analysis. (see below).
[0288] DSC Analysis (See FIG. 3.13)
[0289] .sup.1H NMR (400 MHz, Methanol-d.sub.4)*.delta. 6.80 (s,
1H), 6.75 (s, 1H), 6.09 (d, J=9.6 Hz, 1H), 5.96 (s, 1H), 5.94 (s,
1H), 5.33 (s, 1H), 4.17 (d, J=9.7 Hz, 1H), 3.81 (s, 3H), 3.54 (s,
3H), 2.79 (d, J=15.4 Hz, 2H), 2.71 (d, J=15.4 Hz, 2+1H), 2.23 (d,
J=16.2 Hz, 1H), 1.95 (d, J=16.1 Hz, 1H), 1.49-1.17 (m, 6H), 1.15
(s, 6H). *Partial presuppression of water signal using `watergate`
irradiation
[0290] .sup.13C NMR APT*(101 MHz, Methanol-d.sub.4) .delta. 179.22,
174.90, 174.22, 171.61, 165.22, 149.83, 148.81, 130.80, 126.75,
114.89, 111.84, 102.89, 95.97, 78.30, 76.09, 74.33, 74.01, 71.29,
59.05, 54.23, 53.21, 52.07, 48.95, 44.76, 44.06, 40.88, 40.44,
29.22, 29.18, 29.16, 19.92, 19.10. *APT=Attached Proton Test
[0291] IR (Diamond ATR, solid) cm.sup.-1 2959, 1757, 1732, 1715,
1651, 1580, 1508, 1489, 1464, 1432, 1371, 1305, 1262, 1224, 1186,
1151, 1111, 1081, 1032, 985, 944, 922, 909, 864, 829, 806, 705,
690, 614, 581, 563, 510, 486. See FIG. 1.13
[0292] IR (Diamond ATR, film) cm.sup.-1 3442, 2967, 1738, 1654,
1585, 1505, 1489, 1440, 1373, 1264, 1223, 1115, 1083, 1033, 928.
See FIG. 1.13
[0293] X-Ray Crystallographic Studies
[0294] Single Crystal X-Ray Diffraction (See FIGS. 2.11.1 and
2.11.2)
[0295] From a suspension in its mother liquor, a suitable single
crystal of size 0.58.times.0.36.times.0.28 mm was finally selected
and implemented on the diffractometer.
TABLE-US-00018 Structural data Empirical formula
C.sub.36H.sub.51NO.sub.17 Extended formula
C.sub.29H.sub.40NO.sub.9, C.sub.6H.sub.7O.sub.7, CH.sub.4O Formula
weight 769.78 Temperature 150(2) K Wavelength 0.71073 .ANG. Crystal
system, space group orthorhombic, P 2.sub.1 2.sub.1 2.sub.1 Unit
cell dimensions a = 9.9967(3) .ANG., .alpha. = 90.degree. b =
18.8971(5) .ANG., .beta. = 90.degree. c = 19.2826(7) .ANG., .gamma.
= 90.degree. Volume 3642.6(2) .ANG..sup.3 Z, Calculated density 4,
1.404 (g cm.sup.-1) Absorption coefficient 0.112 mm.sup.-1 F(000)
1640 Crystal size 0.58 .times. 0.36 .times. 0.28 mm Crystal color
colourless Theta range for data collection 2.94 to 27.48.degree.
h_min, h_max -12, 12 k_min, k_max -20, 24 l_min, l_max -25, 13
Reflections collected/unique 18037/4637 [.sup.aR(int) = 0.0424]
Reflections [I > 2.sigma.] 4165 Completeness to theta_max 0.998
Absorption correction type multi-scan Max. and min. transmission
0.969, 0.858 Refinement method Full-matrix least-squares on F.sup.2
Data/restraints/parameters 4637/0/513 .sup.bGoodness-of-fit 1.032
Final R indices [I > 2.sigma.] .sup.cR.sub.1 = 0.0367,
.sup.dwR.sub.2 = 0.0851 R indices (all data) .sup.cR.sub.1 =
0.0427, .sup.dwR.sub.2 = 0.0884 Largest diff. peak and hole 0.289
and -0.214 e .ANG..sup.-3
Atomic coordinates, site occupancy (%) and equivalent isotropic
displacement parameters (A.sup.2.times.10.sup.3). U(eq) is defined
as one third of the trace of the orthogonalized Uij tensor.
[0296] Atom numbering of FIG. 2.11.1 corresponds to below
table.
TABLE-US-00019 Atom x y z occ. U(eq) C1 0.3491(2) 0.92951(11)
0.25699(11) 1 0.0149(4) H1 0.3259 0.9482 0.2129 1 0.018 C2
0.3729(2) 0.96852(11) 0.31322(11) 1 0.0152(4) C3 0.4025(2)
0.92636(11) 0.37724(10) 1 0.0138(4) H3 0.4934 0.9384 0.3953 1 0.017
C4 0.3993(2) 0.84827(11) 0.35162(11) 1 0.0119(4) H4 0.4942 0.8317
0.3535 1 0.014 C5 0.3638(2) 0.85222(11) 0.27257(11) 1 0.0136(4) C6
0.4667(2) 0.81449(12) 0.22685(11) 1 0.0176(5) H6A 0.4901 0.7677
0.2464 1 0.021 H6B 0.5492 0.8431 0.2224 1 0.021 C7 0.3972(3)
0.80632(13) 0.15620(12) 1 0.0220(5) H7A 0.4181 0.8469 0.1256 1
0.026 H7B 0.4258 0.7621 0.133 1 0.026 C8 0.2475(3) 0.80407(12)
0.17319(11) 1 0.0212(5) H8A 0.2 0.8437 0.1504 1 0.025 H8B 0.2075
0.7589 0.1575 1 0.025 N9 0.2393(2) 0.81069(9) 0.25106(9) 1
0.0141(4) HN9 0.251(4) 0.7629(18) 0.2669(17) 1 0.05 C10 0.1057(2)
0.83634(12) 0.27463(11) 1 0.0172(5) H10A 0.0365 0.8014 0.2612 1
0.021 H10B 0.0848 0.8815 0.251 1 0.021 C11 0.1006(2) 0.84755(11)
0.35335(11) 1 0.0144(4) H11A 0.1348 0.8955 0.364 1 0.017 H11B
0.0062 0.8455 0.3686 1 0.017 C12 0.1806(2) 0.79388(11) 0.39455(11)
1 0.0130(4) C13 0.3209(2) 0.79563(10) 0.39455(11) 1 0.0118(4) C14
0.3947(2) 0.74640(11) 0.43291(11) 1 0.0141(4) H14 0.4897 0.7472
0.433 1 0.017 C15 0.3252(2) 0.69701(11) 0.47031(11) 1 0.0153(4) C16
0.1875(2) 0.69386(11) 0.46951(11) 1 0.0153(4) C17 0.1124(2)
0.74148(11) 0.43253(11) 1 0.0159(4) H17 0.0174 0.7392 0.4325 1
0.019 C18 0.2626(2) 0.59964(11) 0.52687(12) 1 0.0212(5) H18A 0.2671
0.5574 0.4966 1 0.025 H18B 0.2624 0.5837 0.5758 1 0.025 C19
0.3571(3) 1.07978(12) 0.25888(13) 1 0.0270(6) H19A 0.2638 1.0737
0.2445 1 0.041 H19B 0.4165 1.0631 0.2219 1 0.041 H19C 0.3746 1.13
0.2679 1 0.041 C21 0.3353(2) 0.98104(10) 0.48356(11) 1 0.0126(4)
C22 0.2160(2) 0.99292(11) 0.53183(11) 1 0.0140(4) C23 0.1479(2)
0.92348(11) 0.55222(11) 1 0.0162(4) H23A 0.0747 0.9336 0.5854 1
0.019 H23B 0.1079 0.9015 0.5105 1 0.019 C24 0.2446(2) 0.87236(11)
0.58470(11) 1 0.0160(4) C25 0.2645(3) 0.77137(12) 0.65633(13) 1
0.0259(5) H25A 0.3417 0.7932 0.6792 1 0.039 H25B 0.2957 0.7397
0.6195 1 0.039 H25C 0.2129 0.7442 0.6904 1 0.039 C31 0.1157(2)
1.04230(11) 0.49544(12) 1 0.0172(5) H31A 0.0748 1.0168 0.4559 1
0.021 H31B 0.0433 1.0545 0.5284 1 0.021 C32 0.1792(2) 1.11061(11)
0.46873(12) 1 0.0195(5) H32A 0.2259 1.1006 0.4245 1 0.023 H32B
0.247 1.1269 0.5026 1 0.023 C33 0.0774(2) 1.16973(11) 0.45699(13) 1
0.0185(5) 33A 0.0362 1.1819 0.5021 1 0.022 H33B 0.0055 1.1515
0.4265 1 0.022 C34 0.1343(2) 1.23774(11) 0.42451(12) 1 0.0177(5)
C35 0.1490(3) 1.23004(14) 0.34612(13) 1 0.0286(6) H35A 0.1813
1.2747 0.3264 1 0.043 H35B 0.062 1.2182 0.3257 1 0.043 H35C 0.2132
1.1923 0.3358 1 0.043 C36 0.2649(3) 1.26062(13) 0.45770(15) 1
0.0279(6) H36A 0.2555 1.26 0.5083 1 0.042 H36B 0.2874 1.3086 0.4423
1 0.042 H36C 0.3363 1.228 0.4439 1 0.042 O1 0.38145(18) 1.03942(8)
0.32111(8) 1 0.0215(4) O2 0.14401(17) 0.63966(8) 0.51222(8) 1
0.0216(4) O3 0.37524(16) 0.64530(8) 0.51409(8) 1 0.0188(3) O4
0.30170(15) 0.93996(7) 0.42992(8) 1 0.0145(3) O5 0.44249(16)
1.00917(8) 0.49151(8) 1 0.0177(3) O6 0.26234(18) 1.02640(8)
0.59345(8) 1 0.0185(3) HO6 0.345(4) 1.0383(18) 0.5906(18) 1 0.05 O7
0.36240(17) 0.87121(9) 0.57473(9) 1 0.0269(4) O8 0.18077(17)
0.82599(8) 0.62673(8) 1 0.0204(4) O9 0.03435(17) 1.29295(8)
0.43436(9) 1 0.0194(4) HO9 0.039(4) 1.3068(17) 0.4747(18) 1 0.047
C51 0.2081(2) 0.62031(12) 0.26814(12) 1 0.0197(5) C52 0.2431(2)
0.55356(11) 0.31079(11) 1 0.0165(4) C53 0.1183(2) 0.53099(11)
0.35211(12) 1 0.0196(5) H53A 0.1 0.5667 0.3885 1 0.023 H53B 0.0402
0.5298 0.3205 1 0.023 C54 0.1343(2) 0.45913(12) 0.38583(12) 1
0.0215(5) O51 0.2755(2) 0.67426(8) 0.28230(9) 1 0.0293(4) O52
0.11948(19) 0.61575(9) 0.22240(10) 1 0.0300(4) O53 0.34983(17)
0.57012(9) 0.35660(9) 1 0.0221(4) HO53 0.358(4) 0.6160(18)
0.3535(18) 1 0.05 O54 0.2279(2) 0.44156(10) 0.42120(12) 1 0.0433(6)
O55 0.03274(18) 0.41626(9) 0.37134(8) 1 0.0204(4) HO55 0.044(4)
0.3790(18) 0.3902(18) 1 0.05 C60 0.2959(2) 0.49411(12) 0.26327(12)
1 0.0204(5) H60A 0.3642 0.5153 0.2324 1 0.025 H60B 0.3427 0.4594
0.2931 1 0.025 C61 0.1992(3) 0.45307(13) 0.21778(13) 1 0.0251(5)
O62 0.0989(2) 0.48721(10) 0.18875(10) 1 0.0344(5) H062 0.104(3)
0.5364(18) 0.2012(18) 1 0.05 O63 0.2167(2) 0.39075(9) 0.20666(11) 1
0.0381(5) O71 0.5434(2) 1.13984(10) 0.43673(10) 1 0.0321(4) HO71
0.498(4) 1.1011(18) 0.4527(18) 1 0.05 C72 0.6645(3) 1.11097(17)
0.41010(17) 1 0.0443(8) H72A 0.6436 1.0742 0.376 1 0.066 H72B
0.7165 1.0903 0.4481 1 0.066 H72C 0.7167 1.1485 0.3879 1 0.066
[0297] X-Ray Powder Diffraction
[0298] The powder sample is well crystallised, with a peak width of
0.127.degree. (2.theta.) at 18.255.degree. (2.theta.). The powder
is constituted in major part by the expected sample referenced
HOCIT 5776. However, the powder pattern reveals the presence of a
second phase, with significant lines at 7.001.degree. (2.theta.)
and 12.317.degree. (2.theta.) for example, not calculated from the
structure determined with a single crystal (for view of diagrams
and experimental details, see FIG. 2.11.3).
Example 15: Preparation and Analyses of Homoharringtonine
Salicylate
##STR00021##
[0300] This ionic compound was obtained from commercial
homoharringtonine mixed with commercial salicylic acid according to
the general procedure in which the solvent was methanol, then
isolated as a white prismatic solid mp 148.7-151.3.degree. C.
(measured by DSC, see FIG. 3.14). Several potentially acceptable
crystals were kept suspended in their mother liquors for the
subsequent X-ray diffraction analysis. (see below).
[0301] DSC Analysis (See FIG. 3.14)
[0302] .sup.1H NMR (400 MHz, Methanol-d.sub.4)*.delta. 7.80 (dd,
J=7.7, 1.7 Hz, 1H), 7.26 (ddd, J=8.8, 7.2, 1.8 Hz, 1H), 6.80-6.70
(m, 4H), 6.09 (d, J=9.6 Hz, 1H), 5.92 (d, J=1.0 Hz, 1H), 5.88 (d,
J=1.0 Hz, 1H), 5.33 (s, 1H), 4.17 (d, J=9.6 Hz, 1H), 3.81 (s, 3H),
3.54 (s, 3H), 3.18 (dd, J=11.0, 6.9 Hz, 1H), 2.71-2.62 (m, 1H),
2.28-2.08 (m, 4H), 1.95 (d, J=16.1 Hz, 1H), 1.47-1.30 (m, 5H),
1.30-1.18 (m, 1H), 1.16 (s, 6H). *Partial presuppression of water
signal using `watergate` irradiation.
[0303] .sup.13C NMR (101 MHz, MeOD)** .delta. 133.50, 131.36,
118.66, 116.85, 114.49, 111.46, 102.48, 95.80, 74.04, 58.71, 53.86,
52.96, 51.78, 49.56, 48.62, 44.45, 43.75, 40.58, 40.24, 28.96,
28.94, 28.86, 19.65, 18.79. **DEPT135: Distortionless Enhancement
by Polarization Transfer (non-quaternary carbons only)
[0304] IR (Diamond ATR, solid) cm.sup.-1 2961.4, 2622.5, 1760.5,
1748, 1740.7, 1722.8, 1651.8, 1625.2, 1590.4, 1579.2, 1503.9,
1487.7, 1459.3, 1374, 1334.4, 1293.2, 1224.3, 1167.4, 1082.6,
1043.9, 1030.5, 995.4, 924.5, 890.7, 857.3, 832.8, 805.3, 763.6,
704.8, 666.2, 613.2, 565.6. See FIG. 1.14
[0305] IR (Diamond ATR, film) cm.sup.-1 3416.8, 2962.9, 2377.4,
2156.9, 1746.7, 1655.2, 1628.2, 1591.3, 1504.8, 1488.2, 1459.5,
1375.8, 1330.2, 1223.6, 1084.2, 1034.5, 930.1, 858.3, 807.3, 763.1,
705.4. See FIG. 1.14
[0306] X-Ray Crystallographic Studies
[0307] Single Crystal X-Ray Diffraction (See FIG. 2.12.1 to
2.12.4)
[0308] From a suspension in its mother liquor, a small single
crystal of size 0.15.times.0.11.times.0.04 mm was finally selected
and implemented on the diffractometer.
TABLE-US-00020 Structural data Empirical formula
C.sub.72H.sub.94N.sub.2O.sub.26 Extended formula
2(C.sub.29H.sub.40NO.sub.9), 2(C.sub.7H.sub.5O.sub.3), 2(H.sub.2O)
Formula weight 1403.5 Temperature 150(2) K Wavelength 0.71073 .ANG.
Crystal system, space group monoclinic, P 2.sub.1 Unit cell
dimensions a = 11.6871(3) .ANG., .alpha. = 90.degree. b =
25.8294(6) .ANG., .beta. = 114.6320(10).degree. c = 12.6300(3)
.ANG., .gamma. = 90.degree. Volume 3465.69(15) .ANG..sup.3 Z,
Calculated density 2, 1.345 (g cm.sup.-1) Absorption coefficient
0.102 mm.sup.-1 F(000) 1496 Crystal size 0.15 .times. 0.11 .times.
0.04 mm Crystal color colourless Theta range for data collection
2.96 to 27.48.degree. h_min, h_max -15, 15 k_min, k_max -33, 33
l_min, l_max -11, 16 Reflections collected/unique 29505/8078
[.sup.aR(int) = 0.0621] Reflections [I > 2.sigma.] 6299
Completeness to theta_max 0.994 Absorption correction type
multi-scan Max. and min. transmission 0.996, 0.886 Refinement
method: Full-matrix least-squares on F.sup.2
Data/restraints/parameters 8078/1/928 .sup.bGoodness-of-fit 1.076
Final R indices [I > 2.sigma.] .sup.cR.sub.1 = 0.0619,
.sup.dwR.sub.2 = 0.121 R indices (all data) .sup.cR.sub.1 = 0.086,
.sup.dwR.sub.2 = 0.1312 Largest diff. peak and hole 0.531 and -0.3
e.ANG..sup.-3
[0309] Atomic coordinates, site occupancy (%) and equivalent
isotropic displacement parameters (A.sup.2.times.10.sup.3).
[0310] U(eq) is defined as one third of the trace of the
orthogonalized Uij tensor.
[0311] Atom numbering of FIG. 2.12.1 corresponds to below
table.
TABLE-US-00021 Atom x y z occ. U(eq) C1 -0.3818(4) 0.00999(17)
0.1577(4) 1 0.0196(9) H1 -0.3779 -0.0157 0.1054 1 0.024 C2
-0.3310(4) 0.00487(17) 0.2729(4) 1 0.0193(9) C3 -0.3588(4)
0.04946(17) 0.3333(4) 1 0.0186(9) H3 -0.4173 0.038 0.3682 1 0.022
C4 -0.4266(4) 0.08958(17) 0.2350(4) 1 0.0172(9) H4 -0.5126 0.0945
0.2325 1 0.021 C5 -0.4453(4) 0.06061(18) 0.1208(4) 1 0.0183(9) C6
-0.5832(4) 0.0587(2) 0.0314(4) 1 0.0298(11 H6A -0.627 0.0292 0.0483
1 0.036 H6B -0.6271 0.091 0.0347 1 0.036 C7 -0.5836(6) 0.0525(3)
-0.0867(5) 1 0.0523(19 H7A -0.6431 0.0772 -0.1428 1 0.063 H7B
-0.6082 0.0169 -0.1162 1 0.063 C8 -0.4507(5) 0.0637(2) -0.0699(4) 1
0.0287(11 H8A -0.4066 0.0313 -0.0719 1 0.034 H8B -0.4502 0.0871
-0.1318 1 0.034 N9 -0.3883(4) 0.08909(15) 0.0468(3) 1 0.0179(8) HN9
-0.406(6) 0.119(3) 0.038(6) 1 0.0 C10 -0.2485(4) 0.08986(18)
0.0908(4) 1 0211(10 H10A -0.2248 0.1127 0.0402 1 0.025 H10B -0.2186
0.0545 0.0856 1 0.025 C11 -0.1831(4) 0.10881(17) 0.2165(4) 1
0.0176(9) H11A -0.1748 0.0794 0.2696 1 0.021 H11B -0.0973 0.1207
0.2312 1 0.021 C12 -0.2525(4) 0.15248(17) 0.2444(4) 1 0.0165(9) C13
-0.3674(4) 0.14288(17) 0.2526(4) 1 0.0151(9) C14 -0.4317(4)
0.18343(17) 0.2773(4) 1 0.0167(9) H14 -0.5087 0.1775 0.2838 1 0.0
C15 -0.3799(4) 0.23192(17) 0.2919(4) 1 0.0181(9) C16 -0.2683(4)
0.24154(17) 0.2810(4) 1 0.0179(9) C17 -0.2022(4) 0.20246(17)
0.2595(4) 1 0.0175(9) H17 -0.1243 0.209 0.2551 1 0.021 C18
-0.3495(5) 0.3174(2) 0.2989(6) 1 0.0342(13 H18A -0.3276 0.3447
0.3591 1 0.041 H18B -0.3985 0.3333 0.2218 1 0.041 C19 -0.2286(5)
-0.07362(19) 0.2749(5) 1 0.0287(11 H19A -0.3051 -0.09 0.218 1 0.043
H19B -0.1753 -0.0999 0.329 1 0.043 H19C -0.1826 -0.0574 0.2342 1
0.043 C21 -0.2376(4) 0.07232(16) 0.5324(4) 1 0.0150(9) C22
-0.1193(4) 0.10173(17) 0.6134(4) 1 0.0181(9) C23 -0.1290(4)
0.15962(16) 0.5815(4) 1 0.0210(10 H23A -0.0436 0.1748 0.6184 1
0.025 H23B -0.1574 0.1625 0.4961 1 0.025 C24 -0.2154(4) 0.19169(17)
0.6156(4) 1 0.0191(9) C25 -0.4265(5) 0.2160(2) 0.5670(5) 1
0.0293(12 H25A -0.4225 0.2098 0.645 1 0.044 H25B -0.5115 0.2084
0.5085 1 0.044 H25C -0.4059 0.2523 0.5602 1 0.044 C31 -0.0033(4)
0.07965(17) 0.6007(4) 1 0.0186(9) H31A -0.0119 0.0866 0.5206 1
0.022 H31B 0.0725 0.0983 0.6551 1 0.022 C32 0.0168(4) 0.02207(17)
0.6244(4) 1 0.0219(10 H32A -0.059 0.003 0.5716 1 0.026 H32B 0.0297
0.0148 0.7056 1 0.026 C33 0.1312(4) 0.00353(17) 0.6058(4) 1
0.0203(10 H33A 0.2055 0.0237 0.6578 1 0.024 H33B 0.117 0.0115
0.5246 1 0.024 C34 0.1622(4) -0.05425(18) 0.6277(4) 1 0.0224(10)
C35 0.0526(5) -0.0876(2) 0.5504(5) 1 0.0333(12) H35A 0.0218 -0.0755
0.4698 1 0.05 H35B -0.0153 -0.0853 0.5766 1 0.05 H35C 0.0803
-0.1237 0.555 1 0.05 C36 0.2022(6) -0.0690(2) 0.7544(5) 1
0.0418(14) H36A 0.2289 -0.1053 0.7657 1 0.063 H36B 0.1312 -0.0643
0.7756 1 0.063 H36C 0.2724 -0.0469 0.8038 1 0.063 O1 -0.2627(3)
-0.03428(12) 0.3396(3) 1 0.0245(7) O2 -0.4215(3) 0.27736(13)
0.3215(3) 1 0.0295(8) O3 -0.2371(3) 0.29334(12) 0.3015(3) 1
0.0291(8) O4 -0.2456(3) 0.07096(12) 0.4237(3) 1 0.0193(7) O5
-0.3115(3) 0.05224(13) 0.5638(3) 1 0.0272(8) O6 -0.1014(3)
0.09442(13) 0.7304(3) 1 0.0232(7) HO6 -0.159(6) 0.100(3) 0.736(6) 1
0.05 O7 -0.1806(3) 0.22291(14) 0.6937(3) 1 0.0343(9) O8 -0.3364(3)
0.18247(12) 0.5481(3) 1 0.0231(7) O9 0.2605(3) -0.06667(14)
0.5922(4) 1 0.0349(9) HO9 0.319(6) -0.049(3) 0.633(6) 1 0.05 C41
-0.4061(4) 0.20979(18) -0.0709(4) 1 0.0185(9) C42 -0.4133(4)
0.26758(17) -0.0845(4) 1 0.0190(9) C43 -0.4734(4) 0.29825(18)
-0.0317(4) 1 0.0236(10) C44 -0.4734(5) 0.3520(2) -0.0387(5) 1
0.0348(13) H44 -0.5137 0.3723 -0.0014 1 0.042 C45 -0.4139(5)
0.3755(2) -0.1008(5) 1 0.0361(13) H45 -0.4138 0.4122 -0.106 1 0.043
C46 -0.3542(5) 0.3463(2) -0.1557(5) 1 0.0320(12) H46 -0.3132 0.3627
-0.1977 1 0.038 C47 -0.3558(4) 0.2929(2) -0.1480(4) 1 0.0252(11)
H47 -0.3169 0.2728 -0.1868 1 0.03 O41 -0.3392(3) 0.18505(13)
-0.1087(3) 1 0.0267(8) O42 -0.4676(3) 0.18943(12) -0.0187(3) 1
0.0218(7) O43 -0.5327(4) 0.27633(14) 0.0297(4) 1 0.0331(9) HO43
-0.525(6) 0.249(3) 0.031(6) 1 0.05 C51 0.0847(4) 0.26596(17)
-0.0909(4) 1 0.0198(10) H51 0.0267 0.2505 -0.161 1 0.024 C52
0.1215(4) 0.31517(18) -0.0810(4) 1 0.0200(10) C53 0.2190(4)
0.32915(16) 0.0374(4) 1 0.0164(9) H53 0.3014 0.3357 0.0333 1 0.02
C54 0.2289(4) 0.27955(16) 0.1123(4) 1 0.0136(9) H54 0.3181 0.2674
0.1417 1 0.016 C55 0.1468(4) 0.23828(16) 0.0226(4) 1 0.0142(8) C56
0.2210(4) 0.18952(17) 0.0199(4) 1 0.0199(9) H56A 0.2763 0.1964
-0.0202 1 0.024 H56B 0.2728 0.1769 0.0997 1 0.024 C57 0.1175(4)
0.15039(18) -0.0484(4) 1 0.0243(10) H57A 0.0894 0.1551 -0.1334 1
0.029 H57B 0.148 0.1144 -0.0276 1 0.029 C58 0.0111(4) 0.16215(17)
-0.0123(4) 1 0.0216(10) H58A 0.0026 0.134 0.0371 1 0.026 H58B
-0.0697 0.166 -0.0817 1 0.026 N59 0.0475(3) 0.21301(14) 0.0560(3) 1
0.0161(8) HN59 0.088(5) 0.199(2) 0.139(5) 1 0.05 C60 -0.0649(4)
0.24526(18) 0.0388(4) 1 0.0199(10) H60A -0.1166 0.2267 0.0718 1
0.024 H60B -0.1165 0.2495 -0.0458 1 0.024 C61 -0.0319(4)
0.29898(18) 0.0951(4) 1 0.0188(9) H61A -0.0155 0.3225 0.0412 1
0.023 H61B -0.1057 0.3126 0.1056 1 0.023 C62 0.0809(4) 0.29972(16)
0.2115(4) 1 0.0143(9) C63 0.2020(4) 0.28854(16) 0.2193(4) 1
0.0135(8) C64 0.3049(4) 0.28691(17) 0.3276(4) 1 0.0156(9) H64
0.3865 0.2784 0.3338 1 0.019 C65 0.2853(4) 0.29777(17) 0.4240(4) 1
0.0179(9) C66 0.1677(4) 0.31120(17) 0.4167(4) 1 0.0197(10) C67
0.0638(4) 0.31097(16) 0.3130(4) 1 0.0188(9) H67 -0.0174 0.3182
0.3094 1 0.023 C68 0.3079(5) 0.3226(2) 0.6008(4) 1 0.0320(12) H68A
0.3372 0.3589 0.6181 1 0.038 H68B 0.3258 0.3046 0.6753 1 0.038 C69
-0.0155(6) 0.3400(2) -0.2692(5) 1 0.0428(15) H69A 0.014 0.314
-0.3086 1 0.064 H69B -0.0446 0.3707 -0.3186 1 0.064 H69C -0.0852
0.3256 -0.2545 1 0.064 C71 0.2427(4) 0.41910(16) 0.0877(4) 1
0.0159(9) C72 0.2104(5) 0.45689(17) 0.1661(5) 1 0.0244(11) C73
0.2446(5) 0.4339(2) 0.2868(4) 1 0.0288(11) H73A 0.2371 0.4613
0.3385 1 0.035 H73B 0.1834 0.4063 0.2808 1 0.035 C74 0.3766(5)
0.4115(2) 0.3417(4) 1 0.0286(11) C75 0.5447(5) 0.3850(3) 0.5159(5)
1 0.0448(15) H75A 0.5447 0.3514 0.4801 1 0.067 H75B 0.5651 0.3802
0.5988 1 0.067 H75C 0.6078 0.4076 0.5075 1 0.067 C81 0.0688(5)
0.46846(19) 0.1066(5) 1 0.0297(12) H81A 0.0225 0.4358 0.1018 1
0.036 H81B 0.0477 0.4926 0.1566 1 0.036 C82 0.0219(5) 0.4915(2)
-0.0146(5) 1 0.0379(14) H82A 0.0717 0.477 -0.0546 1 0.045 H82B
0.0364 0.5294 -0.0077 1 0.045 C83 -0.1172(5) 0.4814(2) -0.0889(5) 1
0.0378(13) H83A -0.1271 0.4443 -0.111 1 0.045 H83B -0.1636 0.4874
-0.0399 1 0.045 C84 -0.1788(5) 0.5126(2) -0.1973(5) 1 0.0353(13)
C85 -0.3125(6) 0.4957(3) -0.2704(6) 1 0.0532(17) H85A -0.3601
0.4953 -0.2222 1 0.08 H85B -0.3123 0.4609 -0.3012 1 0.08 H85C
-0.352 0.52 -0.3352 1 0.08 C86 -0.1053(6) 0.5173(3) -0.2700(5) 1
0.0439(15) H86A -0.1535 0.5378 -0.3398 1 0.066 H86B -0.0895 0.4828
-0.2931 1 0.066 H86C -0.0248 0.5345 -0.2246 1 0.066 O51 0.0848(3)
0.35381(13) -0.1615(3) 1 0.0285(8) O52 0.3708(3) 0.29717(14)
0.5395(3) 1 0.0264(8) O53 0.1754(3) 0.32134(13) 0.5276(3) 1
0.0260(7) O54 0.1836(3) 0.37380(11) 0.0855(3) 1 0.0184(7) O55
0.3039(3) 0.42839(12) 0.0351(3) 1 0.0225(7) O56 0.2839(4)
0.50176(14) 0.1739(4) 1 0.0364(9) HO56 0.272(6) 0.531(3) 0.200(6) 1
0.05 O57 0.4356(3) 0.39631(15) 0.2885(3) 1 0.0382(9) O58 0.4187(4)
0.40908(16) 0.4575(3) 1 0.0409(10) O59 -0.1879(4) 0.56408(15)
-0.1531(4) 1 0.0467(10) H59 -0.2274 0.5839 -0.2093 1 0.07 C91
0.2069(5) 0.15694(19) 0.3512(5) 1 0.0274(11) C92 0.2274(4)
0.16464(17) 0.4767(4) 1 0.0223(10) C93 0.1463(3) 0.19621(14)
0.5016(3) 1 0.0268(11) H93 0.0736 0.2099 0.4399 1 0.032 C94
0.1706(3) 0.20812(14) 0.6171(3) 1 0.0321(12) H94 0.1166 0.2309
0.6341 1 0.039 C95 0.2751(5) 0.1862(2) 0.7069(5) 1 0.0376(13) H95
0.2916 0.1937 0.7856 1 0.045 C96 0.3549(5) 0.1536(2) 0.6824(5) 1
0.0340(13) H96 0.4255 0.1386 0.7441 1 0.041 C97 0.3320(5) 0.1430(2)
0.5688(5) 1 0.0286(11) O91 0.1084(3) 0.17337(13) 0.2713(3) 1
0.0265(8) O92 0.2952(4) 0.13363(15) 0.3347(3) 1 0.0370(9) O93
0.4150(3) 0.11174(15) 0.5479(4) 1 0.0363(9) HO93 0.369(6) 0.114(3)
0.462(6) 1 0.05 OW1 0.4458(4) 0.01162(15) 0.6698(4) 1 0.0458(10)
OW2 0.6810(3) 0.11024(14) 0.7538(3) 1 0.0318(8)
Example 16: Purification of Natural Homoharringtonine as Hydrogen
(2R,3R)-Tartaric Salt
[0312] All operations were performed in a sterile isolator using
dedicated or single-use equipment. The reagents were of
pharmacopoeial quality and all the quality control and quality
assurance written procedures were carried out according to the
current good manufacturing practices. Commercial homoharringtonine
base (100 grams) exhibiting at least 97% of purity was dried then
introduced in a dedicated sterile flask equipped with a stirring
and a refrigerant, and flushed by sterile argon, then 1.2 molar
equivalent of (2R,3R)-(+)-tartaric acid (natural version,
pharmacopoeia quality) was introduced. Then 350 mL of anhydrous
methanol was added under reflux until all solid phase disappeared.
A volume of dry methanol was added to move away from the point of
supersaturation. The homogeneous solution was then withdrawn and
directly filtered hot under vacuum on a microporous filter (0.5
micron). After 15 minutes, fine translucent prismatic crystals of
homohamrngtonine hydrogen (2R, 3R) tartrate begin to form. The
stirred suspension is allowed to stand for 12 hours. At this stage
an in process control (CIP) to check the impurity content of the
crystals and mother liquors. The suspension is drained on a filter
funnel (Buchner) and the crystals are dissolved in hot methanol.
The crystallization operation and the corresponding CIPs are
renewed twice. After the last wringing, the crystals were dried
under vacuum at a temperature of 45c, 20 hours, and then packaged.
Final samples are taken to carry out an analysis report in
accordance with the specifications. The impurity content is less
than 0.3% and the purity (HPC) exceeds 99.7% (the current purity of
semi-synthetic batches). In addition to the usual tests including
microbiological testing and endotoxin detection, all batches of
drug substance were subjected to a high resolution NMR analysis and
a control for in vivo toxicity.
* * * * *
References