Title of Invention

TREATMENT OF ERECTILE DYSFUNCTION AND LIBIDO ENHANCEMENT

Abstract A process for the production of a substance or composition for the therapeutic or prophylactic treatment of erectile dysfunction or the enhancement of libido in a male human or animal subject comprises the step of formulating the substance or composition from at least one of plant material and an extract of plant material of at least one plant species of the family Geraniaceae.
Full Text THIS INVENTION relates to the treatment of erectile dysfunction and
to libido enhancement in a male human or animal subject. More particularly, the
invention relates to a substance or composition for such treatment; to the use of
such substance or composition in the manufacture of a medicament or
preparation; to such treatment; to compounds useful for such treatment and to
derivatives of such compounds; and to a method of such treatment.
Broadly, the invention provides a process for the production of a
substance or composition for the treatment, by therapy or prophylaxis, or erectile
dysfunction, and for the enhancement of libido in a male human or animal subject,
the process comprising the step of formulating the substance or composition from
at least one of plant material and an extract of plant material of at least one plant
species of the family Geraniaceae.
The invention also provides the use of a substance or composition in
the manufacture of a medicament or preparation for the therapeutic or prophylactic
treatment of erectile dysfunction or the enhancement of libido in the male human
or animal body, the substance or composition comprising a formulation of at least
one of plant material, and an extract of plant material of at least one plant species
of the family Geraniaceae
Examples of such plants include M. angustifolia, M. alexandraensi,
M. attenuate (syn. biflora), M. brevirostrata, M. belfastensis, M. burkeana (syn.
glanulata, betschuanica), M. depressa, M. deserticola, M. drudeana, M. galpini, M.
glandulosissima, M. glauca, M. grandifolia (syn. Lanceolate), M. heliotropioides, M.
ignea, M. ignorata, M. I'heritieri, M. lanuginose, M. longipes, M. luederitziana, M.
malvaeflora, M. multifida, M. namaensis, M. natalensis, M. nivea, M. obcordata, M.
ovata (syn. emarginata), M. parvifolia, M. rehmii, M. rudatisii, M. senecioides (syn.

Praemorsa), M. senegalensis (syn. trilobata), M. speciosa L, M. spinosa (syn.
lobata, pilosa), M. stricta, M. transvaalensis, and M. umbellata.
The process may comprise formulating the substance or
composition by size reduction into particulate form, e.g. by grinding (which
includes milling) and/or cutting, of the plant material in the form of stems and/or
roots and/or flowers and/or seeds and/or leaves. The grinding and/or cutting may
be of wet plant material, followed optionally by drying thereof, e.g. by oven-drying.
Instead, the grinding and/or cutting may be of dried plant material, the process
optionally including the step of drying the plant material prior to the grinding and/or
cutting. The process may then include packaging the dried ground plant material
in porous bags of the nature of tea bags, or formulating the particulate plant
material, after the size reduction, into granules, tablets, capsules, oral sprays,
creams or lotions. Naturally, the particle size to which the plant material is
reduced will be selected in accordance with suitability thereof for the final intended
product.
Instead, the formulating may comprise extracting the substance or
composition from the plant material by subjecting the plant material, which has
optionally been subjected to a prior size reduction, to solvent extraction using a
suitable solvent, to obtain a solvent extract containing the substance or
composition. Once again, the plant material subjected to the extraction may be
wet, or it may be dried, having been subjected to an optional drying step prior to
the extraction. If desired, after the extraction, the solvent may be removed to
leave a dried extract comprising the substance or composition.
The extract may thus be selected from organic and aqueous
extracts. The extract may be selected from organic extracts produced by
extraction of plant material with an organic solvent selected from the group
consisting of diethyl ether, isopropyl ether, methanol, ethanol, chloroform,
dichloromethane, ethyl acetate, hexane and suitable mixtures of two or more
thereof or mixtures thereof with water and aqueous extracts produced by

extraction of plant material with water. Examples include
methanol/dichloromethane mixtures and methanol/water mixtures, respectively in
volume ratios, for example, of 1:1 and 9:1.
Instead the extract may be an organic extract produced by extraction
of plant material with a supercritical fluid. The supercritical fluid may be
supercritical carbon dioxide.
The solvent extract, whether in the form of a solution or after drying,
for example by evaporation of organic solvents at atmospheric pressure (including
supercritical fluid solvents), at a reduced pressure depending upon ambient
temperature and pressure (organic solvents), or by freeze-drying or spray-drying
(aqueous solvents), may be refined and/or separated into individual constituents
or compounds, using a suitable technique such as solvent/solvent partitioning
and/or chromatographic separation.
The process may, as indicated above, include carrying out the
solvent extraction on the plant material in finely-divided form, after its having been
subjected to size reduction, for the purpose of facilitating subsequent solvent
extraction, for example by wet or dry grinding or milling, to have a particle size of
at most 6000um, preferably at most 5000um.
For the purpose of the present invention cold solvent extraction at
room or ambient temperature has been found to be adequate at, say 15-30°C, but
the invention contemplates that the possibility of hot extraction at elevated
temperatures of up to 100°C is not excluded.
The Applicant has carried out the present process in particular on
plants on the species Monosonia angustifolia identified as such by the South
African National Biodiversity Institute (SANBI) in Pretoria, South Africa. Plants of
the genus Monosonia are members of the Geraniaceae family, and M. angustifolia
is a perennial herb with five petalled pink flowers whose geographic distribution is

in open grassland throughout South Africa. A herbarium specimen has been
deposited at the SANBI in Pretoria under Genspec. No. 39250002. Extracts
obtained from Monosonia angustifolia,\r\ particular solvent extracts using a
methanol/dichloromethane mixture as solvent, have been shown to be promising
for the treatment of erectile dysfunction and libido enhancement in male subjects.
Fractionation of organic solvent extracts, such as dichloromethane or
1:1 by volume methanol/dichloromethane solvent extracts for example by
solvent/solvent partitioning followed by column chromatography, has isolated five
compounds believed to be active in the treatment or prophylaxis of erectile
dysfunction and/or libido enhancement. The process may accordingly include
isolating active compounds from the extracts, for example by fractionation such as
by chromatography, e.g. column chromatography, followed by recombining the
active compounds.
The finished product can thus be in the form of a combination of the
active compounds and/or the other chemical ingredients that are present in the
plant extract. The active compounds may be extracted, as indicated above, by
extraction techniques which include aqueous extraction, organic solvent
extraction, and super-critical fluid extraction, or related chemical extraction
methods, followed by drying (removal of the extracting solvent) by spray-drying,
freeze-drying or evaporation. The extract can then be separated by solvent-solvent
partitioning coupled with chromatographic separation techniques to give the active
compounds. The active compounds can be formulated, for example and in
particular, into suitable tablets, capsules and oral sprays.
According to another aspect of the invention, there is provided the
use of at least one compound selected from compounds of the formulae (1) and
(2)




(1) (2)
in which
R1, R2, R4, R5, R6 are independently selected from H and C1-4 alkyl and
R3 is independently selected from H and OC1-4 alkyl, or
R1 and R2 together form CR8R9 in which R8 and R9 are independently
selected from H and C1-4 alkyl and R3-R6 are as defined above, or
R5 and R6 together form CR8R9 in which R8 and R9 are independently
selected from H and C1-4 alkyl and R1-R4 are as defined above, and
A is CH2 and B is CO or
B is CH2 and A is CO, and
R7 is CH2-aryl
in the manufacture of a medicament for the treatment by therapy or prophylaxis of
erectile dysfunction or the enhancement of libido in a male human or animal
subject.
R1, R2 and R4-R6 may be selected from Me and H and R8 and R9
may be H.

The CH2-aryl substituent may be

in which R1 and R2 are as defined above.
In particular, the compounds of formula (1) and (2) may be selected
from
9-(1',3'-benzodioxol-5'-yl)-4,5,6,7-tetramethoxynaphtho[2,3-c]furan-1(3H)-one
(Compound 3)
9-(1',3'-benzodioxol-5'-yl)-4,6,7-trimethoxynaphtho[2,3-c]furan-1(3H)-one
{Compound 4)
9-(3',4'-dimethoxyphenyl)-4-hydroxy-6,7-methylenedioxynaphtho[2,3-c]furan-
1 (3H)-one (Compound 5)
3-(1,,3'-benzodioxol-5'-ylmethylene)-4-(3",4"-dimethoxybenzyl)dihydrofuran-2(δH)-
one (Compound 6)
4-(3',4'-dimethoxyphenyl)-9-methoxy-6,7-methylenedioxynaphtho[2,3-c]furan-
1 (3H)-one (Compound 7)
These compounds are also called 5-methoxyjusticidin A (compound
3), justicidin A (compound 4), chinensinaphthol (compound 5), suchilactone
(compound 6) and retrochinensinaphthol methyl ether (compound 7).
The invention extends to a pharmaceutical composition for the
therapeutic or prophylactic treatment of erectile dysfunction or the enhancement of
libido in the male human or animal body, the composition comprising a formulation

of at least one of plant material, and an extract of plant material of at least one
plant species of the family Geraniaceae.
The plant material and the extract may be as hereinbefore described.
The invention extends to a pharmaceutical composition for the
treatment by therapy or prophylaxis of erectile dysfunction or the enhancement of
libido in a male human or animal subject, the composition including at least one
compound selected from compounds of formula (1) and (2).




(1) (2)
in which
R1, R2, R4, R5, R6 are independently selected from H and C1-4 alkyl and
R3 is independently selected from H and OC1-4 alkyl, or
R1 and R2 together form CR8R9 in which R8 andR9 are independently
selected from H and C1-4 alkyl and R3-R6 are as defined above, or
R5 and R6 together form CR8R9 in which R8 and R9 are independently
selected from H and C1-4 alkyl and R1-R4 are as defined above, and
A is CH2 and B is CO or
B is CH2 and A is CO, and
R7 is CH2-aryl.

R1, R2 and R4-R6 may be selected from Me and H and R8 and R9
may be H.
The CH2-aryl substituent may be

in which R1 and R2 are as defined in any one of Claims 1 to 3 inclusive.
The compounds of formula (1) and (2) may be selected from
compounds (3)-(7).
The structures of compounds 3-7, are set out below



The elucidation of the structures of the isolated Components 3 - 7,
which were isolated by column chromatography from a sample obtained from the
plant material of Monsonia angustifolia by solvent extraction using a 1:1 by volume
material/dichloromethane mixture, followed by drying, partitioning between a 90:10
methanol/water mixture and hexane, then partitioning the methanol/water layer so
obtained with dichloromethane and evaporating the dichloromethane layer, was
based essentially on spectral analysis. The 1H and 13C NMR (nuclear magnetic
resonance) spectral data provided the first stage in the characterization of the
compounds. When necessary, advanced homonuclear and heteronuclear two-
dimensional NMR methods such as COSY (proton correlation spectroscopy),
HMQC (heteronuclear multiple quantum correlation) and HMBC (heteronuclear
multiple bond correlation) were applied to achieve the complete assignments of

the 1H and 13C correlations of the isolated compounds. From the proton decoupled
DEPT (distortionless enhancement by polarization transfer) pulse sequence sub-
spectra, protonated carbons could be assigned according to their multiplicity. The
resulting 13C and 1H chemical shift experimental data of the isolated active
Compounds 3 - 7 is collated in Tables 1, 2, 3, 4 and 5 hereunder.
To confirm the so-established structures, the chemical shift data
were compared to the data of the previously identified and known compounds on-
line or using structural databases. The structures of the five compounds were
elucidated as aryl naphthalene lignans which are biosynthetically formed by fusing
at least two phenyl propanyl (C6-C3) units. Compound 3 was unambiguously
identified as previously isolated from the plant Justicia procumbens and is known
as 5-Methoxyjusticidin A (Siani, Antonio C et al Journal of Natural Products, 61(6),
796-797, 1998) with a molecular formula C23H20O8and a corresponding molecular
ion at m/z 424[M]+. Compound 4 has been identified as Justicidin A with m/z 394
[M]+ (molecular formula C22H18O7). This compound is one of the most commonly
known lignans found in plants (M. Okigawa et al, Tetrahedron, 26, 4301 - 4305,
1970, L. Badheka et al, Phytochemistry, 25(2), 487 - 489 1986). Compound 5, is
commonly known as Chinensinaphthol and it was previously isolated from Justicia
procumbens (Z. Horii et al Chemical Communications, 653, 1968). A mass charge
of m/z 380 was obtained for this compound, which mass charge is in agreement
with the proposed molecular formula of C21H1607. Compound 6 (molecular
formula C21H20O6) was found to have m/z 368 [M]+ and it was assigned the name
Suchilactone based on literature and NMR spectral data comparison with the
authenticated published data (S. Ghosal et al, Phytochemistry, 13, 1933, 1974,
Biswanath Das and Ratna Das, Phytochemistry, 40(3), 931 - 932, 1995).
Compound 7 (molecular formula C22H18O7) was elucidated as
Retrochinesinaphthol methyl ether in conjunction with spectral data (Z. Horii et al,
Chemical and Pharmaceutical Bulletin, 19(3), 535 - 537, 1971) and a molecular
ion [M]+ of m/z 394 was obtained.

The invention extends to the use of a substance or composition in
the manufacture of a medicament or preparation for the therapeutic or prophylactic
treatment of erectile dysfunction and for libido enhancement, in the male human or
animal body, the substance or composition comprising a formulation obtained from
plant material from a plant of the genus Monsonia, in particular Monsonia
angustifolia.
The invention also extends to the use of a substance or composition
in the manufacture of a medicament or preparation for the therapeutic or
prophylactic treatment of erectile dysfunction and for libido enhancement in the
male human or animal body, the substance or composition comprising at least one
compound selected from the group consisting of Compound 3, Compound 4,
Compound 5, Compound 6 and Compound 7 as identified above.
The substance or composition may comprise a plurality of said
Compounds 3, 4, 5, 6 and 7; and for it may comprise one or more derivatives of
said Compounds 3, 4, 5, 6, and 7.
The invention also extends to substances and compositions for the
treatment, by therapy and prophylaxis, of erectile dysfunction and for libido
enhancement in the male human or animal body, the substances or compositions
comprising formulations, in particular solvent extracts, (and the active compounds
contained therein), obtained from plant material from plants of the genus
Monsonia.
More particularly, the invention extends to a substance or
composition for use in a method of treatment of erectile dysfunction and for libido
enhancement, in the male human or animal body, by administering an effective
amount of said substance or composition to the human or animal body, said
substance of composition forming part of a formulation, in particular a plant
extract, obtained from plant material of a plant which is a member of the genus
Monsonia.

The substance or composition may comprise at least one of
Compounds 3 - 7 identified above or at least one derivative thereof. Preferably the
substance or composition comprises all of Compounds 3 - 7 identified above and,
optionally, one or more derivatives thereof.
The invention extends further to a substance or composition for use
in a method of treatment by therapy or prophylaxis of erectile dysfunction and for
libido enhancement, in the male human or animal body, by administering an
effective amount of said substance or composition to the human or animal body,
the substance or composition comprising at least one of Compounds 3 - 7
identified above.
The substance or composition may comprise a plurality of, and
preferably all of, said Compounds 3-7, and it may comprise one or more
derivatives thereof.
The one or more of Compounds 3 - 7, or the derivatives thereof, may
be present in the substance or composition at a total concentration or proportion of
at least 5 % by mass, preferably at least 10 % by mass and more preferably at
least 20 % by mass.
In particular the substance or composition may be in a form selected
from particles of the plant material which has been subjected to size reduction and
contained optionally in porous bags of the nature of tea bags, or in the form of
tablets, capsules, oral sprays, syrups and possibly herbal tinctures. Thus, the
substance or composition may be for use in a method of treatment or prophylaxis,
or a method of enhancement, in which the substance or composition is
administered in unit dosage form. Preferably, the administering of the substance
or composition is such as to attain an effective total serum concentration by mass
of said one or more of Compounds 3 - 7 identified above, or one or more
derivatives thereof, in the human or animal body. More particularly, the

administration of the substance or composition may be at an effective daily dosage
rate, the rate being dependent on the body mass of the subject.
The invention extends further to novel derivatives of any one of
Compounds 3 - 7 identified above which can be obtained from an extract of a plant
of the genus Monsonia, in particular Monsonia angustifolia.
The invention furthermore extends to a method of treatment by
therapy or prophylaxis of erectile dysfunction and to the enhancement of libido, in
the male human or animal body, the method comprising administering to a male
human or animal subject an effective amount of a substance or composition
comprising an extract from a plant of the genus Monsonia, in particular M.
angustifolia.
The invention furthermore extends also to a method of treatment by
therapy or prophylaxis of erectile dysfunction and to libido enhancement, in the
male human or animal body, the method comprising administering to a male
human or animal subject an effective amount of a substance or composition
comprising at least one of Compounds 3 - 7 identified above.
The substance or composition may comprise a plurality, preferably
all, of said Compounds 3 - 7.
The administering of said substance or composition may be in unit
dosage form. In particular, the administering of said substance or composition
may be such as to attain an effective total serum concentration by mass or one or
more of said Compounds 3 - 5, in the human or animal body. More particularly,
the substance or composition may be administered at an effective daily dosage
rate, the rate being dependent on the body mass of the subject.
As described in somewhat more detail hereunder, the present
invention provides a plant-derived treatment for alleviating erectile dysfunction and

for enhancing libido in a male subject. The substances or compositions of the
present invention have been demonstrated to be effective in enhancing the libido
of male rats in terms of a so-called mating frequency/pregnancy model (MFM).
From this enhancement of libido it can be inferred that the substances and
compositions of the present invention will also be effective in alleviating erectile
dysfunction in a male subject.
The invention will now be described, by way of non-limiting
illustrative example, with reference to the following worked Examples:
EXAMPLE 1
Extracts were prepared of plant material from the plant species
M.angustifolia, identified as such by the SANBI in Pretoria. The plant material
comprised the roots, stems, leaves and purple flowers, from which plant material
the extracts were prepared.
Initial liquid-liquid partitioning of the extract afforded hexane, dichloromethane and
water fractions. Based on PDE 5 inhibition, the dichloromethane fraction showed a
20% improvement in activity compared to the original crude extract. Repeated
column chromatography on flash silica gel afforded the isolation of five
compounds that were identified as aryl naphthalene lignans 3 to 7, which are
formed biosynthetically from two phenyl propanyl (C6-C3) units.
Characterization of 5-methoxyjusticidin A (3)
5-Methoxyjusticidin A (3) was isolated as a white amorphous solid. A molecular
ion peak at m/z 425.4 corresponding to [M+1]+ in the positive LRESIMS and at m/z
424 in AutoSpec ETOF El+ which are both pointing to the molecular formula
C23H20O8. The UV spectrum was obtained from a diode array detector (DAD) and
absorptions were observed at A 365, 263 and 237 nm.

In the 1H NMR spectra of 3 (Table 1) the presence of a methylenedioxy group, ΔH
6.02 (1H, d) and δH 6.07 (1H, d) with a very small coupling of J 1.3 Hz was
observed. The other methylene singlet observed at δH 5.41 is characteristic of the
methylene group found in a penta-lactone ring of lignans. Four singlets assigned
to the methoxyl groups were observed at δH 3.74, 3.95, 3.97 and 4.01. An
aromatic proton singlet was observed for H-8 at SH 6.94. Three aromatic protons
were found exhibiting the ortho coupling (J = 7.9 Hz) and meta coupling (J = 1.6
Hz) which are consistent with a 1,3,4 trisubstituted phenyl group.
The 13C NMR spectra showed 23 resonance signals. Four signals could
immediately be assigned to the four methoxyl carbons resonating at δc 55.8, 61.4,
62.4, and 62.0. The multiplicity of the other carbon signals was obtained from
DEPT spectra that showed 10 protonated carbon signals, two of them being CH2
and four were CH, apart from the four that were already assigned as methoxy
groups. There was also evidence of a carbonyl group at δc 169.5, which is
characteristic of the lactone functionality in the strained five-membered ring. C-3
was also not found in the aromatic region and was assigned to the methylene
group at δc 66.5 attached to the carbonyl forming a lactone. The structure was
confirmed by the >1 JCH correlations observed in a HMBC experiment.
5-Methoxyjusticidin A 3 was previously isolated by Siani et al. from the wood of
Protium unifoliolatum1. In this article, some of the NMR signals were not assigned
correctly (C-9, C-9a, C-8, and C-8a) and we were able to assign them with the aid
of 2D experiments. The author acknowledged the wrong assignment and thus the
assignment we propose can be taken as correct.


Characterization of justicidin A (4)
This compound was isolated as white amorphous flakes. The 1H NMR spectrum
of 4 was similar to that of 3. The main difference was that the spectrum of 4
exhibits the presence of only three methoxy groups (δH 3.79, 4.04, 4.11) instead
of four as for 3. One methoxy at C-5 was replaced by a proton resonating at δH
7.52. The absence of observed coupling between the two protons resonating at δH
7.52 and 7.04 suggests a para relationship between them. The methylene signal
at ΔH 5.51 which forms part of the furanone ring was still present as was the three
aromatic protons that formed the 1, 3, 4-trisubstituted phenyl group. A pair of

proton doublets with very weak coupling (δH 6.02 and 6.06 with a JHH value of 0.9
Hz) was observed for the methylenedioxy substituent.
Twenty two 13C NMR signals were observed which is in compliance with one less
methoxy group. The three methoxy groups were assigned at δc 55.8, 56.1 and
59.7. A carbonyl signal was observed downfield at δc 169.5. The methylene group
forming part of the furanone ring was assigned at δc 66.6. The methylenedioxy
carbon signal was found at δc 101.2. The rest of the carbon signals were found to
support the structural framework with the same substitution pattern as for 5-
methoxyjusticidin A (3).
Justicidin A (4) showed a molecular ion peak of m/z 395.4 in ESIMS positive
mode corresponding to [M+1]+ and the molecular formula C22H18O7. In the UV
spectrum of this compound as obtained from a diode array detector has a strong
absorption maximum at 261.7 nm. This compound is widely distributed within the
different species of Justicia and it has been widely studied before for antiviral" ,
anti platelet"'™ and cytotoxic activity but not for the activity reported here.
Published NMR data for justicidin AVI,V" supported the structural assignment of
justicidin A (4).


Characterization of chinensinaphthol (5)
Compound 5 is commonly known as chinensinaphthol and it was previously
isolated from Justicia procumbens and tested for antiplatelet properties as
documented by Z. Horii et al 1968.viii
A mass to charge ratio (m/z) of 381.4 observed in LRESIMS in positive mode was
assigned to the molecular ion peak [M+1]+, which is in agreement with the formula
C21H16O7. The UV maximum plot of this compound from the DAD spectrum
showed absorptions at 322.0, 266.4 and 228.8 nm, which is characteristic of aryl
naphthalene molecules.

Compound 5 was purified by flash silica gel chromatography and visualization
under long range UV 254 nm gave a purple coloured spot and a very intense blue
fluorescent color when viewed at 366 nm. It was obtained as a white amorphous
substance that could not be re-dissolved in a single solvent only. Thus, it was
dissolved in a mixture of CDCI3 and MeOH-d4 for NMR experiments and the
obtained data are collated in Table 6.3.
1H NMR data showed a singlet at ΔH 5.35 which can be assigned to the lactone
methylene group and a resonance at δH 6.15 (s, H-10) that is characteristic of a
methylenedioxy group, and two methoxyl groups appeared as singlets at ΔH 3.71
and 3.84. In the previous two structures (3 and 4), the methylenedioxy groups
were attached to the C ring and the methoxy groups to the A ring. However,>1 JCH
correlations observed in the HMBC experiment, indicated that in this compound,
the methylene group is on the A ring and the methoxy substitutents on the B ring.
The three aromatic C ring protons formed an ABX system characteristic of
trisubstituted phenyl as it was observed for the lignans 3 and 4. These protons
resonated at δH 6.83 (H-2\ d, 8.24), 6.77 (H-6\ dd, 2.07 & 8.03), 7.05 (H-5\ d,
8.24) and their coupling was supported by the COSY 2D correlations.


13C NMR spectra data with 21 carbons supported the proposed structure 5 and
the mass spectrum data obtained. The carbonyl C-1 of the lactone ring and the
methylene group C-3 in the same ring were assigned resonances at δc 169.4 and
66.4 respectively. Other important signals were the two methoxy groups that
resonated at δc 55.4 and 55.5 and the methylenedioxy C-10 that resonated at δc
101.9.
On the basis of the spectral data and the supporting correlations observed in 2D
NMR experiments, compound 3 was identified as chinensinaphthol.

Characterization of suchilactone, 6
Crystalline and yellowish 6 was isolated from the methanol-dichloromethane
extract of M. angustifolia and its spectral data is collated in Table 6.4. The
structure was deduced with the help of the 2D NMR correlations that provided
information to the C-H connectivity pattern. The proposed structure was also
confirmed by comparison to literature data lx
ESIMS (+) of compound 6 showed a molecular ion peak of m/z 368 which agrees
with a molecular formula of C21H20O6. In the mass spectrum, fragments were
observed at m/z 151, which was assigned to the fragment 6.1 and at m/z 217
assigned to the fragment 6.2.
Two methoxy groups were observed in the 1H NMR spectrum at ΔH 3.84 and 3.86
as singlets. In contrast to the other compounds where only one 1,3,4-trisubstituted
phenyl ring was observed, in the 1H NMR spectrum of 6, a second set of ABX
protons were present. Additionally, an olefinic proton was observed at δH 7.50.



From the COSY correlations it was observed that the protons at H-4 couples with
those protons at positions H- 5 and H-7". These key correlations are shown in Fig
6.4 and they are represented by arrows in red. The other correlations observed
were H-6' to H-4' and H-7' and also in the C ring there was a strong correlation
between H-5" and H6".
The 13C NMR of compound 6 showed 21 carbon signals with two methoxy groups
overlapping at δc 55.9. The carbonyl in the lactone ring resonates at δc 172.5 and
the methylene group in the same ring appears at δc 69.8. The exocyclic double
bond of the lactone ring showed resonance peaks at δc 126.1 and 137.2 for the
carbons at C-3 and C-8', respectively. The methylene carbon at C-7" was

assigned to the signal at δc 37.7. The methylenedioxy group at C-2' showed its
resonance peak at δc 101.7. The DEPT spectrum was used to assign the
multiplicity of the carbon resonances. Two overlapping methoxy peaks, three CH2
and eight CH peaks, leaving the remaining eight carbon signals to be assigned as
quaternary carbon atoms.
Characterization of retrochinesinaphthol methyl ether, 7
Compound 7 (C22H18O7) was identified as retrochinesinaphthol methyl ether.x A
mass- charge ratio m/z 395.4 [M+1]+ (ESIMS) was obtained which supports the
molecular formula of C22H18O7. The structure of the compound is similar to that of
5 with the difference that the methylene and the carbonyl groups of the lactone
ring are swopped around and that the hydroxyl group in position C-4 was now
replaced by a methoxy group.



Comparison of the 1H and 13C NMR data of this compound (7) with those reported
in the literaturex and observed for 5 led to the assignment of compound 7 as
retrochinesinaphthol methyl ether.
EXAMPLE 2
Plant material from M.angustifolia was oven-dried at 35 - 45° (i.e. 40
± 5°C) to more or less constant mass and then ground, to a particle size of 1 -
2mm, in a hammer mill. A quantity of 408g of dried starting material, after milling,
yielded 352g in the 1 - 2mm particle size range. To 86g of the dried plant
material was added M of de-ionized water to form a mixture which was boiled for
1 hour. The boiled mixture was allowed to cool to room temperature, filtered and
freeze-dried for 24 hours to yield 22.16g of brownish fluffy powder, forming an
aqueous extract.
To 266g of similarly dried and ground plant material was added 6t of
a 1:1 by volume methanol/dichloromethane mixture, to form a solvent/plant
material mixture which was allowed to stand at room temperature for 1 hour with
occasional stirring. The solvent/plant material mixture was filtered and the
residual plant material was again similarly extracted using a said 1:1 by volume
methanol/dichloromethane mixture for 1 hour with occasional stirring. The two
filtrates so obtained were combined and evaporated under vacuum at a sub-
atmospheric pressure depending upon atmospheric temperature and moisture
content to leave 27.32g of residue, in the form of a sticky dark-green extract.
An amount of 1.5g of the sticky dark-green methanol/dichloromethane
extract was partitioned into two layers between M of a 9:1 by volume
methanol/water mixture and 500ml of hexane. The hexane layer was separated

and the methanol /water mixture evaporated at 40°C to obtain 395.8mg of a
sticky greenish fraction, to which water was added up to a total volume of 500ml,
which was then further extracted using a 500ml batch of dichloromethane. This
dichloromethane batch was separated from the water and the water was then
further extracted using a further 500ml batch of dichloromethane which was also
separated from the water, and the water was then finally extracted with a final
500ml batch of dichloromethane which was separated therefrom. The three
batches of dichloromethane separated from the water were combined and
evaporated at 40°C to yield 542.6mg of a brownish extract, while the separated
residual water was freeze-dried to yield 473.8mg of a brownish powder.
A quantity of 6g of the sticky dark-green diethyl methanol/dichloromethane
extract was fractionated and purified using flash chromatography (silica gel 60,
Merck, 230 - 400 mesh) and a 1.5:8.5 by volume ethyl acetate/hexane mixture as
eluant to produce 163.6mg of a steroid P-sitosterol and the Compounds 3-7
identified above, which were lignan compounds and which were respectively
produced in quantities of 35 mg (Compound 3), 19.3 mg (Compound 4), 27.2 mg
(Compound 5), 36.4 mg (Compound 6) and 16.5 mg (Compound 7).
Methods were tested for the manufacture of a finished product in a
form which is suitable for use by humans and intended to be used for the
manufacture of a finished dose form. Two methods were used to prepare the
products wherein, in one method, fresh wet plant roots, stems, leaves and flowers
were cut, boiled in water, filtered and dried to a powder by either spray-drying or
freeze-drying. Alternatively the cut plant roots, stems, leaves and flowers were
oven-dried by using an oven set to dry at 35 - 45°C (i.e. 40 ± 5°C or by using an
herb dryer. The dried plant parts were ground to granules having a particle size of
at most 6000pm and were thereafter extracted by hot water at about 97°C,
filtered and dried to a powder using spray-drying or freeze-drying techniques.
In the extraction of the dried plant material 16.00kg of the stems,
leaves and flowers was cut to 5mm lengths and dried in an herb drier using the

drying cycle set forth in Table 6 hereunder. The dry (brownish-green) material
was ground to fine granules (3.38 kg having a particle size of at most 6000um)
and extracted by boiling in 75l of water for an hour. The water was filtered and
the residual plant material was boiled in an additional 75l water for 30 minutes.
The combined filtered water extract (115l) was left to stand overnight in a cold
room. An amount of 500ml of the water extract was freeze-dried for 24 hours to
produce 40.6g of a dry brownish powder. The remaining 114.5l was spray-dried
(see Table 7 hereunder for the spray-drying conditions employed) and produced
804g of dry brownish powder which appeared substantially similar to the 40.6g
freeze-dried sample, both visually and chemically. These two extracts were
compared by HPLC-MS and TLC methods, which showed them to be similar to
each other as well as to the 22.1g of brownish fluffy powder produced by freeze-
drying the aqueous extract described initially.
Table 6: Drying parameters for the herb dryer

In the extraction of the fresh wet plant material a combination of the
fresh plant stems, leaves and flowers (5.65 kg) was sliced to 5mm lengths and
boiled in 40l of water for an hour. The water was filtered and the residual plant
material was boiled in additional 40l of water for 30 minutes. The extracts so
obtained were combined and filtered to yield 46.62l of a brownish aqueous extract
in the form of a sap. The sap was left in a cold room overnight. Spray-drying (see
Table 5 hereunder for spray drying conditions) of the sap generated 242g of a
brownish powder. The powder was shown by HPLC-MS and TLC methods to be
similar to the 22.16g of brownish fluffy powder produced by freeze-drying the
aqueous extract described initially, and to the 804g of brownish powder obtained
by spray-drying the aqueous extract of dried plant material described above.

Table 7: Spray drying parameters

A 6.00g portion of the 804g of powder which was prepared by spray-
drying the aqueous extract of dried plant material as described above was
formulated into twenty effervescent tablets at a dose of 300 mg of the spray-dried
powder/tablet (see Table 8 hereunder for formulation compositions).
Table 8: Ingredients used in the formulation of effervescence tablets

A further 6.00g portion of the 804g of powder which was prepared by
spray-drying the aqueous extract of dried plant material as described above was
formulated into twenty capsules each containing 300 mg of the spray-dried
powder. The capsules were produced and contained the ingredients as set forth in
Table 9 hereunder.

Table 9: Ingredients used in Capsules formulations

To ascertain the efficacy of the invention, the aqueous and organic
extracts described above were tested. A mixture of part of the 22.16g of brownish
fluffy powder initial aqueous extract and part of the 27.32g of sticky dark-green
methanol/dichloromethane extract, prior to the partitioning, was formed by mixing
in a ratio of 4:6 by mass and was tested according to the available in vivo sexual
activity, mating frequency/pregnancy model (MFM). The model is a measure of
male libido. The extract mixture was administered orally to 3 male rats at a dose of
300mg/kg for 8 consecutive days. Sexual activity was recorded by video
surveillance during the nocturnal period i.e. from 17h00 to midnight. The vehicle
control substance used was distilled water dosed orally to each animal at a daily
dose of 10ml/kg.
The extract mixture exhibited a significant increase in mating
frequency and the number of pregnant female rats as compared to the vehicle
control, thereby demonstrating increased male libido. The results obtained from
the bioassay are shown in Tables 10 and 11 hereunder.

Table 10: Results obtained from the Sexual Activity, Mating frequency
Assay
Table 11: Results obtained from the Sexual Activity, Pregnancy Assay


In order to establish the possible mode of action of the prepared
extracts for the MFM activity observed in rats, the extracts were also screened in
vitro for inhibition of the phosphodiesterase 5 enzyme. Inhibition of the
phosphodiesterase 5 enzyme limits the hydrolysis of cyclic Guanisine
Monophosphate (cGMP) to acyclic Guanisine Monophosphate (5'GMP) and
thereby increases the intracellular concentrations of cGMP which results in the
relaxation of the corpus carvenosum smooth muscle. When this muscle is relaxed
blood flows and fills the arteries causing the penis to erect.
The aqueous extract and the methanol/dichloromethane extract
whose 4:6 by mass combination was the test substance in Tables 6 and 7 above
were formed into a different combination by mixing in a 1:1 ratio by mass, which
1:1 combination was tested for the in vitro inhibition of the phosphodiesterase 5
enzyme. It exhibited a significant (80%) inhibition at 100μg/mf (see Table 12
hereunder).

Further in vitro assaying of part of said 27.32g of sticky dark-green
methanol/dichloromethane extract and of part of the 22.16g of brownish fluffy
powder aqueous extract was carried out separately in the phosphodiesterase 5
enzyme inhibition assay to establish which extract contributed most to the activity
of the 1:1 combination. The methane/dichloromethane extract exhibited the
highest inhibition of the phosphodiesterase 5 enzyme (see Table 12). It is also
worth noting that the enzyme inhibition that was observed from the 1:1 by mass
combination was approximately the average of the inhibition observed from the
respective methanol/dichloromethane and aqueous extracts and it is measured
significant (ie. the results are considered to be significant if more than 50%
inhibition is measured).
The assay of the inhibition of the phosphodiesterase 5 enzyme was
carried out by the MDS Pharma Laboratory in Taiwan. Phosphodiesterase 5
enzyme, partially purified from human platelets, was used. Test compounds
and/or vehicles were incubated with 3.5 μg enzyme and 1 μM cGMP containing
0.01 μM [3H]cGMP in Tris buffer at pH 7.5 for 20 minutes at 30°C. The reaction
was terminated by boiling for 2 minutes and the resulting GMP was converted to
guanosine by addition of 10 mg/ml snake venom nucleotidase and further
incubation at 30°C for 10 minutes. Unhydrolyzed cGMP was bound to AG1-X2
resin, and remaining [3H]guanosine in the aqueous phase was quantitated by
scintillation counting. Compounds were screened at 100 ΜM. Since enzyme
activity could change from lot to lot, the concentration used was adjusted if
necessary.
In order to isolate and identify the active ingredients responsible for
the inhibitory activity against the phosphodiesterase 5 enzyme, further
fractionation through liquid-liquid partitioning was conducted on part of the 27.32g
of sticky dark-green methanol/dichloromethane extract. The dichloromethane,
hexane and water fractions obtained therefrom by the partitioning between
methanol/water and hexane and further dichloromethane extraction described

above were screened for in vitro phosphodiesterase 5 enzyme inhibition. Only the
dichloromethane fraction resulted in a substantial increase in the inhibition of
phosphodiesterase 5 enzyme relative to the crude plant extracts.
Table 12: Summary of results demonstrating inhibition

As the inhibition of the phosphodiesterase 5 enzyme is one of the
mode of actions that may facilitate the relaxation of the corpus carvenosum
smooth muscle, part of the 22.16g of the brownish fluffy powder aqueous extract
and part of the 27.32g of sticky dark-green methanol/dichloromethane extract
were also tested for the relaxation of the pre-contracted rabbit corpus carvernosal
smooth muscle.
Measurement of rabbit corpus cavernosum smooth muscle
relaxation/contraction was assayed by the University of Pretoria Assay
Laboratory in Pretoria, South Africa in accordance with the bioassay as described
by Levin et al. 1997 Journal of Andrology, Volume 18, No. 3, pages 246 - 249
(with some minor changes). Strips (12mm long and 1-2mm thick) of rabbit corpus
carvernosal smooth muscle were dissected and mounted in an organ-bath
chamber containing Krebs-PSS solution with the following composition: NaCI =
7.01 g/l, KCI = 0.34g/l, KH2P04 = 0.1 g/l, NaHC03 = 1.99g/l, CaCI2 = 0.2g/l,
MgS04 = 0.3g/l and glucose = 1.8g/t One end of the muscle was tied to the
inside bottom of the perfusion bath and the other end to a thin wire connected to
a Harvard isotonic force transducer for isotonic tension measurements. Changes
in isotonic tension were recorded on a computerized calibrating program. The

corpus cavernosum muscle was perfused with 2ml Krebs-PSS buffered saline
solution and oxygenated with 95% 02 and 5% C02 for 5 minutes to establish a
stable base-line recording. This was followed by perfusion with 2ml of high CaCI2
in Krebs-PSS (17.8 mg/ml) for muscle contraction. Base-line tension was set at
the point of maximal contraction following the addition of CaCI2 into the
experimental bath. The extracts to be analyzed were added after a stable
contraction base-line. The same procedure was repeated for the positive control,
Sildenafil. The contraction/relaxation was reported relative to Sildenafil tested at
78 ng/ml. In these experiments the stimulation frequency used for rabbit strips
was 9 Hz. Both of these extracts demonstrated a potential to relax the rabbit
corpus carvenosum smooth muscle with the aqueous extract demonstrating this
potential more significantly than the methanol/dichloromethane extract (see Table
13 hereunder).
Table 13: Results obtained from the assaying of extracts in the rabbit corpus
carvernosal smooth muscle assay.

Although the aqueous extract showed more significant relaxation of
the smooth muscle as compared to the methanol/dichloromethane extract, the
difference was not conclusive. However the latter exhibited a higher
phosphodiesterase 5 enzyme inhibition than the former (see Table 12 above).
The significant phosphodiesterase 5 enzyme activity shown by the
methanol/dichloromethane extract necessitated further investigation, by
chromatographic purification thereof in order to isolate the components
responsible for the observed activity.

Fractionation of the methanol/dichloromethane extract resulted in the
isolation of Compounds 3-7, all of which were shown to be lignans as illustrated
above. The five lignans were also shown to be present in the dichloromethane
fraction using thin layer chromatography (TLC) and HPLC methods. These lignans
were screened for phosphodiesterase enzyme inhibition as described above by
MDS Pharma, they were screened as described hereunder for the relaxation of the
pre-contracted rabbit smooth muscle and they were screened as described
hereunder for cytotoxicity against the Chinese hamster ovarian cells (CHO) the
results of which are summarized in Table 14 hereunder.
Rabbit corpus cavemosum relaxation/contraction was measured by
the MDS Pharma Laboratory. Corpus cavemosum obtained from New Zealand-
derived albino male rabbits weighing 2.5-3 kg and sacrificed by CO2 overexposure
was used. A strip from the basal area of the corpus cavernosal was removed and
placed under 2g tension in a 10 ml bath containing Krebs solution at pH 7.4 at
32°C and sub-maximal isometrically recorded tonic contraction was induced by
phenylephrine (3 uM). The test substance (30uM) induced relaxation by 50% or
more (>50%) within 5 minutes, relative to the control 0.3uM sodium nitroprusside
response, indicating significant relaxation.
In vitro cytotoxicity against mammalian cell line was assayed against
Chinese hamster ovarian cells by the Assay Laboratory of the Department of
Medicine, Division of Pharmacology, University of Cape Town, South Africa.
Samples of Compounds 3-7 were tested for in vitro cytotoxicity
against the Chinese Hamster Ovarian cell line using the 3-(4,5-dimethylthiazol-2-
yl)-2,5-diphenyltetrazoliumbromide (MTT) assay. All samples were tested in
triplicate on a single occasion. The MTT-assay was used as a colorimetric assay
for cellular growth and survival, and compares well with other available assays
(Mosman, Journal of Immunological Methods 65, 55-63 (1983) and Rubinstein et
al., Journal of the National Cancer Institute 82, 1113-8 (1990)). The tetrazolium
salt MTT was used to measure all growth and chemo-ensitivity.

Aqueous samples were dissolved in water and organic samples were
dissolved in a methanol: water mixture (1:9) by volume. The initial concentration of
stock solutions was 2mg/ml for all samples. Samples were stored at -20°C until
use. The highest concentration of methanol to which the cells were exposed to
had no measurable effect on the cell viability (data not shown). Emetine was used
as the positive control in all experiments. The initial concentration of emetine was
100ug/ml, which was serially diluted in complete medium with 10-fold dilutions to
give 6 concentrations, the lowest being 0.001 μg/mt. The same dilution technique
was applied to all other samples with an initial concentration of 100μg/ml to give 5
concentrations, with the lowest concentration being 0.01 μg/ml.
The 50% inhibitory concentration (IC5o) values for these samples
were obtained from dose-response curves, using a non-linear dose-response
curve fitting analyses via GraphPad Prism v 2.01 software.
Table 14: Bioassay results for Compounds 3 to 7

Phosphodiesterase 5 enzyme inhibition and smooth muscle relaxation were considered significant
at >50% values IC50 values tests, Emetine was used as the control with an IC50 value of 0 04μg/ml
Viagra showed 100% relaxation at 18X10~2μg/ml or an IC50 value of 4 1X10'3μM for
phosphodiesterase 5 enzyme inhibition
Significant inhibition was obtained for Compound 7 in the
phosphodiesterase 5 enzymatic assay. Rabbit corpus cavernosum smooth muscle
relaxation was observed for Compounds 3, 5, 6, and 7. The results indicate that,

while these compounds demonstrated significant relaxation of the rabbit corpus
cavernosum smooth muscle, their ability to inhibit the phosphodiesterase 5
enzyme was limited (except for Compound 7). This indicates that the Compounds
3, 5, 6 and 7 could act through a different mode of action. In addition the whole
plant extract may act by inhibiting the phosphodiesterase 5 enzyme through a
synergistic effect.
The isolated lignan compounds were also tested for cytotoxicity
against the Chinese hamster ovarian cells (Table 14 above) and Emetine was
used as the control with an IC5o value of 0.04μg/ml (values ≤1.0 are considered to
be cytotoxic). The four Compounds 3, 5, 6, and 7 that have relaxation effect on
the pre-contracted rabbit smooth muscle are non-cytotoxic at the test
concentrations with the least toxic being Compounds 6 and 7. The cytotoxic
Compound 3 according to this Chinese hamster ovarian cell test, did not exhibit
any relaxation of the rabbit smooth muscle and demonstrated an insignificant
inhibition of the phosphodiesterase 5 enzyme at the Compounds' test
concentrations. Thus Compounds 3, 5, 6 and 7 have a therapeutic window of
interest based on the evaluated concentrations.
EXAMPLE 3
Extracts of M.angustifolia were compared with extracts of M.galpinii
and M. brevirostrata to show the presence of compounds (3) - (7) in these plants.
The results are shown in Table 15.
Method of extract preparation:
The dried plant material was ground and successively extracted with methanol-
dichloromethane (1:1, v/v). The solvent was filtered and evaporated to dryness in
a rotary evaporator in vacuo. The generated organic extracts were stored in a cold
room at -20 °C when not in use.

HPLC analysis:
Sample preparation:
Samples were reconstituted in a 50:50 methanol: acetonitrile mixture, vortexed
for 30 seconds, filtered and injected.
Romil methanol and acetonitrile were used throughout.
HPLC method:
Waters 2695 HPLC Pump:
Solvent gradient:



Table 15

The present invention can, at least potentially, be regarded as having
the following beneficial properties:
a) A treatment for the erectile dysfunction as well as for increasing male libido
is provided;
b) Extracts of the plant that induce the relaxation of the pre-contracted corpus
cavernosum smooth muscle demonstrate a potential treatment for erectile
dysfunction;
c) Compounds 3, 5, 6, and 7 isolated from the extracts of the plant
demonstrate a potential for the treatment of erectile dysfunction through
relaxation of the pre-contracted corpus cavernosum smooth muscle without
inhibition of the phosphodiesterase 5 enzyme, thereby reducing any unwanted
side effects which may occur through the inhibition of this enzyme; and
d) The extracts and compounds for the treatment of both erectile dysfunction
and for increasing male libido.

References
i. Siani, A. C, Zoghbi, M. das G. B., Wolter, E.L.A. and Vencato, I.,
1998. J. Nat. Prod, 61, 796-797.
ii. Asano, J., Chiba, K., Tada, M., and Yoshii, T., 1996. Phytochemistry,
42,713-717.
iii. Gonzalez, A.G., Perez, T.P. and Trujillo, J.M., 1978. Tetrahedron,
34, 1011-1013.
iv Chen, C, Hsin, W., Ko, F., Huang, Y., Ou, J. and Teng, C, 1996, J.
Nat. Prod. 59, 1149-1150.
v. Fukamiya, N. and Lee, K., 1986. J. Nat. Prod. 49, 348-350.
vi. Okigawa, M., Maeda, T. and Kawano, N., 1970. Tetrahedron, 26,
4301-4305.
vii. Badheka, LP., Prabhu, B.R. and Mulchandani, N.B., 1986.
Phytochemistry, 25, 487-489.
viii. Horri, Z., Ohkawa, K., Kim, S. and Momose, T., 1968. Chem.
Commun., 653-655.
ix. Ghosal, S., Chauhan, R.P.S. and Srivastava, R.S., 1974.
Phytochemistry, 13, 1933-1936.
x. Horii, Z., Ohkawa, K., Kim, S.W. and Momose, T. 1971. Chem.
Pharm. Bull., 19, 535-537.

WE CLAIM :
1. A process for the production of a substance or composition for the
therapeutic or prophylactic treatment of erectile dysfunction or the enhancement of
libido in a male human or animal subject, the process comprising the step of
formulating the substance or composition from at least one of plant material and
an extract of plant material of at least one plant species of the Genus Monsonia,
the material and the extract including at least one compound selected from
9-(1,3'-benzodioxol-5'-yl)-4,5,6,7-tetramethoxynaphtho[2,3-c]furan-1(3H)-one
(Compound 3)
9-(3',4'-dimethoxyphenyl)-4-hydroxy-6,7-methylenedioxynaphtho[2,3-c]furan-
1 (3H)-one (Compound 5)
3-(1',3'-benzodioxol-5'-ylmethylene)-4-(3",4"-dimethoxybenzyl)dihydrofuran-2(δH)-
one (Compound 6)
4-(3',4,-dimethoxyphenyl)-9-methoxy-6,7-methylenedioxynaphtho[2,3-c]furan-
1 (3H)-one (Compound 7).
2. A process for the production of a substance or composition for the
therapeutic or prophylactic treatment of erectile dysfunction or the enhancement of
libido in a male human or animal subject, the process comprising the step of
formulating the substance or composition from at least one of plant material and
an extract of plant material in which the plant is M. angustifolia.
3. A process as claimed in Claim 1 or Claim 2, in which the extract is selected
from organic and aqueous extracts.
4. A process as claimed in Claim 3, in which the extract is selected from
organic extracts produced by extraction of plant material with an organic solvent
selected from the group consisting of diethyl ether, isopropyl ether, methanol,
ethanol, chloroform, dichloromethane, ethyl acetate, hexane and suitable mixtures

of two or more thereof or mixtures thereof with water and aqueous extracts
produced by extraction of plant material with water.
5. A process as claimed in Claim 3, in which the extract is an extract produced
by extraction of plant material with a supercritical fluid.
6. A process as claimed in Claim 5, in which the supercritical fluid is
supercritical carbon dioxide.
7. A pharmaceutical composition for the therapeutic or prophylactic treatment
of erectile dysfunction or the enhancement of libido in the male human or animal
body, the composition comprising a formulation of at least one of plant material,
and an extract of plant material of at least one plant species of the Genus
Monsonia, the material and the extract including at least one compound selected
from 9-(1 ',3'-benzodioxol-5'-yl)-4,5,6,7-tetramethoxynaphtho[2,3-c]furan-1 (3H)-
one (Compound 3)
9-(3',4'-dimethoxyphenyl)-4-hydroxy-6,7-methylenedioxynaphtho[2,3-c]furan-
1(3H)-one (Compound 5)
3-(1',3,-benzodioxol-5*-ylmethylene)-4-(3",4"-dimethoxybenzyl)dihydrofuran-2(δH)-
one (Compound 6)
4-(3',4'-dimethoxyphenyl)-9-methoxy-6,7-methylenedioxynaphtho[2,3-c]furan-
1(3H)-one (Compound 7).
8. A pharmaceutical composition for the therapeutic or prophylactic treatment
of erectile dysfunction or the enhancement of libido in the male human or animal
body, the composition comprising a formulation of at least one of plant material,
and an extract of plant material, in which the plant species is M. angustifolia.
9. A pharmaceutical composition as claimed in Claim 7 or Claim 8, in which
the extract is selected from organic and aqueous extracts.

10. A pharmaceutical composition as claimed in Claim 9, in which the extract is
selected from organic extracts produced by extraction of plant material with an
organic solvent selected from the group consisting of diethyl ether, isopropyl ether,
methanol, ethanol, chloroform, dichloromethane, ethyl acetate, hexane and
suitable mixtures of two or more thereof or mixtures thereof with water and
aqueous extracts produced by extraction of plant material with water.
11. A pharmaceutical composition as claimed in Claim 9, in which the extract is
an organic extract produced by extraction of plant material with a supercritical
fluid.
12. A pharmaceutical composition as claimed in Claim 11, in which the
supercritical fluid is supercritical carbon dioxide.
13. A pharmaceutical composition for the treatment by therapy or prophylaxis
of erectile dysfunction or the enhancement of libido in a male human or animal
subject, the composition including at least one compound selected from
9-(1',3'-benzodioxol-5'-yl)-4,5,6,7-tetramethoxynaphtho[2,3-c]furan-1(3H)-one
(Compound 3)
9-(3',4'-dimethoxyphenyl)-4-hydroxy-6,7-methylenedioxynaphtho[2,3-c]furan-
1(3H)-one (Compound 5)
3-(1',3'-benzodioxol-5'-ylmethylene)-4-(3",4"-dimethoxybenzyl)dihydrofuran-2(5/-/)-
one (Compound 6)
4-(3',4'-dimethoxyphenyl)-9-methoxy-6,7-methylenedioxynaphtho[2,3-c]furan-
1(3H)-one (Compound 7).

A process for the production of a substance or composition for the therapeutic or prophylactic treatment of erectile dysfunction or the enhancement of libido in a male human or animal subject comprises the step of formulating the substance or composition from at least one of plant material and an extract of plant material of at least one plant species of the family Geraniaceae.

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Patent Number 265328
Indian Patent Application Number 5212/KOLNP/2008
PG Journal Number 08/2015
Publication Date 20-Feb-2015
Grant Date 19-Feb-2015
Date of Filing 22-Dec-2008
Name of Patentee CSIR
Applicant Address SCIENTIA, 0002 PRETORIA
Inventors:
# Inventor's Name Inventor's Address
1 KHOROMBI, ERIC 41 RONELDA PARK, 456 BUSH ROAD, WILLOW PARK MANOR, PRETORIA, 0002
2 FOUCHE, GERDA 485 MULBERRY STREET, MORELETA PARK, PRETORIA, 0044
3 MAHARAJ, VINESH JAICHAND 25 NAALDEHOUT STREET, HEUWELOORD, CENTURION, PRETORIA, 0149
PCT International Classification Number A61K 31/365
PCT International Application Number PCT/IB2007/051951
PCT International Filing date 2007-05-23
PCT Conventions:
# PCT Application Number Date of Convention Priority Country
1 2006/04334 2006-05-25 South Africa