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A Randomized, Double-Blind, Placebo-Controlled Trial to Determine the Effectiveness of Botanically Derived Inhibitors of 5- ? -Reductase in the Treatment of Androgenetic Alopecia

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Androgenetic alopecia (AGA) is characterized by the structural miniaturization of androgen-sensitive hair follicles in susceptible individuals and is anatomically defined within a given pattern of the scalp. Biochemically, one contributing factor of this disorder is the conversion of testosterone (T) to dihydrotestosterone (DHT) via the enzyme 5-alpha reductase (5AR). This metabolism is also key to the onset and progression of benign prostatic hyperplasia (BPH). Furthermore, AGA has also been shown to be responsive to drugs and agents used to treat BPH. Of note, certain botanical compounds have previously demonstrated efficacy against BPH. Here, we report the first example of a placebo-controlled, double-blind study undertaken in order to examine the benefit of these botanical substances in the treatment of AGA. The goal of this study was to test botanically derived 5AR inhibitors, specifically the liposterolic extract of Serenoa repens (LSESr) and beta-sitosterol, in the treatment of AGA. Subjects: Included in this study were males between the ages of 23 and 64 years of age, in good health, with mild to moderate AGA. The results of this pilot study showed a highly positive response to treatment. The blinded investigative staff assessment report showed that 60% of (6/10) study subjects dosed with the active study formulation were rated as improved at the final visit. This study establishes the effectiveness of naturally occurring 5AR inhibitors against AGA for the first time, and justifies the expansion to larger trials.
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A Randomized, Double-Blind, Placebo-Controlled Trial to Determine the
Effectiveness of Botanically Derived Inhibitors of 5AR in the Treatment of
Androgenetic Alopecia
Nelson Prager*, Karen Bickett*, Nita French, and Geno Marcovici†
*Clinical Research and Development Network, Aurora, CO; French and Associates,
Atlanta, GA; †Advanced Restoration Technologies, Denver CO.
Running Title:
Botanicals in the Treatment of AGA
Keywords:
androgenetic alopecia, clinical trial, saw palmetto, ß-sitosterol, male pattern
baldness
Address for Correspondence:
Geno Marcovici, Ph.D.
Chief Scientific Officer
Advanced Restoration Technologies, Inc.
Denver, CO
Email: dr.marcovici@hairgenesis.com
Abstract
Background
Androgenetic alopecia (AGA) is characterized by the structural miniaturization of
androgen-sensitive hair follicles in susceptible individuals and anatomically defined
within a given pattern of the scalp. Biochemically, one contributing factor of this
disorder is the endogenous conversion of testosterone (T) to dihydrotestosterone (DHT)
2
via the enzyme 5-alpha reductase (5AR). This metabolism is also key to the onset and
progression of benign prostatic hyperplasia (BPH). Further, AGA has also been shown
to be responsive to drugs and agents used to treat BPH. Of note, certain botanical
compounds have previously demonstrated efficacy against BPH. Here, we report the
first example of a placebo-controlled, double-blind study undertaken in order to examine
the benefit of these botanical substances in the treatment of AGA.
Objectives
The goal of this study was to test botanically derived 5AR inhibitors, specifically
the liposterolic extract of Serenoa repens (LSESr) and ß-sitosterol, in the treatment of
AGA.
Subjects
Included in this study were males between the ages of 23 and 64 years of age, in
good health, with mild to moderate AGA.
Results
The results of this pilot study showed a highly positive response to treatment.
The blinded investigative staff assessment report showed that 60% (6/10) study subjects
dosed with the active study formulation were rated as improved at the final visit.
Conclusions
This study establishes the effectiveness of naturally occurring 5AR inhibitors
against AGA for the first time, and justifies the expansion to larger trials.
Introduction
Androgenetic alopecia (AGA) shares a number of endocrinolgic pathways with
benign prostatic hyperplasia (BPH). Certain botanical compounds, and specifically those
under investigation herein, have previously demonstrated the ability to inhibit key
metabolic processes associated with BPH. Based on the parallel etiologies of these two
disorders, it was the central hypothesis of this pilot research study to examine the
putative benefit these botanicals may offer in the treatment of AGA.
Androgenetic alopecia (AGA) is characterized by a receding hairline and/or hair
loss within a specific pattern on the scalp (Shapiro et al. 2000). This condition, which
affects both men and women, is inherited as a polygenic disorder likely involving several
genes and multiple pathways, although the precise mechanism(s) remain unknown.
However, one factor, which has been demonstrated to contribute to the pathogenesis of
3
this condition, involves a genetically predetermined sensitivity to the effects of the
androgenic hormone dihydrotestosterone, or DHT, in certain scalp hair follicles
(Mowszowicz et al. 1993). DHT is believed to shorten the growth, or anagen, phase of
the hair cycle, causing miniaturization of the follicles, and producing progressively finer
hairs.
The production of DHT (reduced from T) is catalyzed by the enzyme 5-alpha
reductase (5AR). In the prostate gland and in susceptible scalp hair follicles, androgen
responsive cells express the genes encoding the steroid enzyme 5AR. 5AR is a
membrane-bound enzyme that catalyzes the irreversible conversion of T to DHT (Itami
et al. 1994). Two isozymes exist: the type 1 enzyme, encoded by the SRD5A1 gene,
localized to chromosome 5p15, and the type 2 isozyme, encoded by the SRD5A2 gene
localized to chromosome 2p23 (Morissette et al. 1996). Immunolocalization studies
have shown that the type 1 enzyme is expressed primarily in newborn scalp, and in skin
and liver, and the type 2 isozyme protein is expressed primarily in genital skin, liver and
the prostate (Negri-Cesi et al. 1999). In the prostate gland, the conversion of T to DHT
by 5AR has been strongly implicated in the pathogenesis of BPH. Of note, the
endocrine dysfunction associated with BPH bears a striking similarity to that linked to
AGA.
BPH affects almost all men to some degree as they age, and can cause a
significant disruption of lifestyle due to urinary outflow obstruction and irritative
symptoms. BPH is characterized clinically by large, discrete nodules formed in the
periurethral region of the prostate. These nodules may narrow the urethra sufficiently to
cause full or partial obstruction. An accumulation of estrogen in the aging prostate,
along with increased conversion of testosterone to its more active metabolite,
dihydrotestosterone (DHT), has been shown to contribute BPH, however, the specific
etiology of BPH remains unknown. Nonetheless, studies have demonstrated that by
blocking the conversion of T to DHT, with either pharmaceuticals or natural compounds,
the circulating level of DHT is reduced by 80%, the size of the prostate gland is reduced
by about 20% and the level of prostate-specific antigen (PSA) drops by about 50%
(Mikolajczyk et al. 2000).
4
Interestingly, it has also been observed that eunuchs (males who have been
castrated prior to the onset of puberty) do not develop BPH, nor do they develop AGA,
and moreover, after castration BPH has been shown to regress (Wilson et al. 1999).
Since normal testicular function appears to be necessary for the development of BPH, it
is believed that the hyperplastic tissue metabolizes androgenic hormones differently than
normal prostate tissue. Although by definition this tissue is benign, progressive growth of
the tumor may cause significant obstruction of the urethra and interfere with the normal
flow of urine (Wilt et al. 2000b). As with AGA, the incidence of BPH increases with
advancing age. BPH is so common, that it is believed all men will develop benign
prostatic hyperplasia if they live long enough. Some degree of BPH is present in 80%
of all men over 40 years old and this figure increases to 95% of all men 80 years old
(Guthrie and Siegel 1999).
In support of a relationship between AGA and BPH, several lines of evidence
have converged to support the view that both circulating testosterone and the modifying
enzyme, 5AR, have profound effects on androgen metabolism, and provide insight into
the role of these hormones play in hair loss. Specifically, the absence of circulating
testosterone and, therefore, its metabolite DHT, in males castrated prior to puberty has
been shown to prevent AGA in later life. These findings demonstrate the importance of
this metabolism in the pathogenesis of male pattern hair loss (Giltay and Gooren 2000).
Of equal relevance, it has been observed that, line in a group of male
pseudohermaphrodites presenting inherited mutations in the 5AR gene, the preservation
of the juvenile hair line invariably occurs (Imperato-McGinley et al 1990). These two
examples demonstrate that whether the disturbance in androgen metabolism results
from either the absence of substrate (T), the active metabolite DHT, or dysfunction of the
enzyme 5AR, at least one phenotypic consequence is consistent and reproducible -
terminal hair density as well as juvenile hairlines remain intact in such individuals.
In contrast, a third and critical observation has been noted in bodybuilders who
self-administer anabolic steroids, in that excessive levels of circulating T and therefore
DHT had the opposite effect of the first two examples – that of acceleration of hair loss in
genetically susceptible individuals due to upregulation of the hormonal processes which
result in AGA (Lise et al 1999).
5
Collectively, these lines of evidence point to a common theme among disorders
resulting from dysregulation of the conversion of T to DHT, and have led to the central
hypothesis of this research. Until now, the only treatments available for AGA have
consisted essentially of the topically administered drug Rogaine™ (minoxidil 2% & 5%)
and the orally delivered pharmaceutical Propecia™ (finasteride 1mg).
Rogaine (topically applied minoxidil 2%--5%) is the best known drug in the
category of medical AGA treatment. Minoxidil delivered via oral ingestion was initially
indicated in the treatment of refractory hypertension. It was noted to cause
hypertrichosis (increased nonsexual hair growth), however, the mechanism by which it
stimulates hair growth remains unknown. Clinical trials have shown that a 2% solution
applied topically to the scalp can stimulate hair growth in some men and women
(Rushton et al. 1989).
Finasteride 1 mg (Propecia™, Merck) was approved by the US FDA December,
1997 for the treatment of male pattern hair loss (androgenetic alopecia, AGA) in men
only. Safety and efficacy were demonstrated in men between 18 and 41 years of age
with mild to moderate hair loss of the vertex and anterior mid-scalp area. Efficacy in bi-
temporal recession has not been established (Kaplan and Olsson, 1996). Propecia™ is
not approved for use in women or children. Finasteride is a preferential, competitive
inhibitor of the intracellular, type II, 5 alpha-reductase isoenzyme which converts
testosterone into dihydrotestosterone (DHT), the more potent androgen. In humans, the
type II 5 alpha-reductase isoenzyme is primarily found in the root sheath of the hair
follicle, prostate, seminal vesicles, epididymis, fetal genital skin and in fibroblasts from
normal adult genital skin, as well as liver, and is responsible for two-thirds of circulating
DHT (Sawaya and Price 1997). In target organs, finasteride treatment is thought to result
in selective androgen deprivation affecting DHT without lowering circulating levels of
testosterone, thus preserving the desired androgen mediated effects on muscle strength,
bone density and sexual function. As noted above, the balding scalp in AGA contains
miniaturized hair follicles and increased amounts of DHT compared with non balding
scalp, and finasteride treatment produces inhibition of the isozyme, resulting in a rapid
reduction in scalp and serum DHT concentrations.
6
Since AGA shares similar hormonal pathways with BPH, it was previously
recognized that the pharmaceutical agents useful against BPH may offer some potential
benefit in the treatment of AGA. The modification of Proscar™ (finasteride 5mg—initially
indicated for BPH), to Propecia™ (finasteride 1mg—new indication AGA) serves as a
paradigm for this rationale (Kaufmann 1999). Like finasteride, several botanically
derived substances have also demonstrated the ability to inhibit key hormonal processes
associated with BPH. Importantly, these botanicals have not been linked with the
spectrum of negative side effects, adverse reactions, or teratogenicity, associated with
the pharmaceutically derived alternatives (Klepser and Klepser, 1999).
Recently, several clinical trials have been reported demonstrating the efficacy of
botanical compounds in the treatment of a number of androgen dependent conditions,
and, specifically, BPH. For example, among 1,098 BPH patients tested in one recent
study, the general safety profile of the lipsterolic extract of Serenoa repens (LSESr 320
mg/day), or saw palmetto berry extract, compared favorably with that of finasteride, and
sexual side effects were less common with the extract than with the drug. In particular
the use of this extract has not been associated with erectile dysfunction, ejaculatory
disturbance, or altered libido (Wilt et al. 2000a). Remarkably, in another biochemical
study, it was found that LSESr was a 3-fold more effective inhibitor than finasteride (5
mg/day) at concentrations adjusted to the recommended doses for BPH treatment. It
should be noted that finasteride as indicated for AGA is dosed significantly lower (1
mg/day), suggesting, a 15-fold more potent level of inhibition at the recommended daily
dose of LSESr (320 mg/day) (Delos et al. 1994).
LSESr is the first line of BPH treatment in Europe and elsewhere, and is believed
to act by inhibiting 5AR. Several in vitro experiments in cultured human foreskin
fibroblasts have found LSESr to be a strong and specific inhibitor of the enzyme 5AR
(Swoboda and Kopp, 1999). In vitro studies have also shown that LSESr inhibits both
isozymes, whereas finasteride selectively inhibits only type 2 5AR (Iehle et al 1995).
Moreover, finasteride demonstrates a solely competitive inhibition, while LSESr has
additionally been found to display noncompetitive as well as uncompetitive inhibition of
the type 1 as well as type 2 5AR isozyme (Gerber 2000).
7
As with LSESr, the primary indication for treatment with the plant phytosterol ß-
sitosterol has been in the treatment of BPH. In addition to a potential role in 5AR
inhibition, as a minor component of LSESr, ß-sitosterol in particular has been shown to
lower the bioavailability of cholesterol in the local environment in animal models, thereby
suggesting it may inhibit downstream steriod hormone biosynthesis, most specifically
testosterone (Wang and Ng, 1999). One potential mechanism of action of ß-sitosterol in
the prostate and the hair follicle, therefore, may involve a local reduction of T (substrate)
in the microenvironment of 5AR active tissues.
In one large clinical study, a total of 519 men undertook a 26 week double-blind
controlled study testing ß-sitosterol against placebo in assessing urinary flow as the
operative criterion. Compared with placebo, ß-sitosterol significantly improved urinary
symptom scores and flow measures (Bracher 1997). Based on the success of ß-
sitosterol in the treatment of BPH, as well as other factors, it was incorporated as an
active component of the AGA trial study formulation. IN RECOGNITION OF THE
EFFICACY THESE BOTANICALLY DERIVED 5AR INHIBITORS DEMONSTRATED IN
THE TREATMENT OF BPH, IT WAS THE CENTRAL HYPOTHESIS OF THIS STUDY
TO TEST THEM FOR THE FIRST TIME AGAINST AGA.
8
Subjects and Methods
Patient Population
INCLUDED IN THIS STUDY WERE
males between the ages of 23 and 64 years
of age, in good health, with mild to moderate AGA. Exclusion criteria CONSISTED OF
1) those who had been on prescription or over-the-counter treatment for AGA or a
prostate condition within the past 30 days prior to initiation of the trial; 2) symptomatic
cardiac problems, uncontrolled hypertension, symptomatic hypotension, autoimmune
disorders; 3) those who had participated in a clinical trial within 30 days prior to
enrollment 4) those with any known allergy to any ingredients in the test products; and 5)
those with any other medical condition that could interfere with successful conclusion of
the study. Subjects enrolled in this study met guidelines established under the
Hamilton/Norwood scale of hair loss. Study subjects presented with moderate to
significant male pattern hair loss in the vertex area (grade II vertex through grade VI
vertex). Baseline characteristics of study subjects were reasonably well matched and fell
within the desired study protocol inclusion parameters. The protocol was approved by
the Western Institutional Review Board, Olympia, SA, and all investigation was
performed following written informed consent in accordance with these guidelines. All
investigative staff members participating in this study were trained to evaluate and report
the parameters described in the study protocol.
Study Protocol
After informed consent, approximately twenty-six (26) male subjects aged 23 to
64 with AGA were initially evaluated and randomly assigned, in a double-blind manner,
to one of the following groups: either active oral softgel supplement, 1 softgel twice daily
or matching oral placebo, 1 softgel twice daily.
The composition of the softgels, a GMP-compliant product manufactured by
Softgel Technologies Inc. Los Angeles CA, encased in an inert carob colored soluble
shell, was as follows: The active softgel components consisted of ß-sitosterol 50mg, and
saw palmetto extract (standardized to 85-95% liposterolic content) 200mg. Systemic
absorption of the active softgel components' bioavailability WAS
enhanced by the use of
9
lecithin 50mg, inositol 100mg, phosphatidyl choline 25mg, niacin 15mg, biotin 100mcg.
The historical experience of other investigators testing these botanicals against BPH
was an important factor in determination of the recommended drug-dosage levels for this
study.
Historically, a number of animal studies support the safety, efficacy, and dosage
for these botanical agents, however, no animal research was undertaken as a part of
this proof-of-concept trial (Schilcher 1999). The placebo soft gels, encased in an inert
carob colored soluble shell, were composed as follows: soybean oil 540mg.
Test product and placebo were prepared by Softgel Technologies Inc. (Los
Angeles, CA) to be identical in appearance. Product was randomly assigned code
numbers. Codes were not available to involved personnel until after completion of the
trial and final data review. Study participation encompassed a duration of approximately
4.6 months, with a maximum duration of approximately 5.4 months. There were three
scheduled clinic visits. Baseline/randomization (approximately week 0), enrollment
(approximately week 0), conclusion (approximately week 21). The enrollment visit (Visit
1) was also the randomization visit. During this visit subjects were randomized, study
medication was dispensed, and baseline investigative staff evaluations were made. The
block size for this study was 30 with a treatment vs. control ratio of 1:1. All subjects
were instructed to use Neutrogena T/Gel or similar shampoo to standardize any variable
which might otherwise occur by the use of differing shampoo formulations.
Written informed consent was obtained from all study subjects prior to entry, with
a copy given to each subject. A brief medical history was obtained including vital signs
(weight, blood pressure, heart rate. Overall hair assessment was made using a
standardized 7 point scale (Table I).
Subjects were asked to rate their current level of satisfaction with their hair.
Subjects were then assigned to treatment in sequential order and given instructions
regarding study agent. Study supplies were then dispensed and subjects were sent
home to begin the study.
10
Study completion
As with the enrollment visit, a brief medical history was obtained including vital
signs (weight, blood pressure, heart rate). Overall hair assessment was made using a
standardized 7 point scale (Table I). Subjects were asked to evaluate any change with
respect to their current level of satisfaction with their hair.
Efficacy Measures
Efficacy measures were administered at baseline, and final visit. These were: 1)
investigative staff assessed hair growth (Figure 1) and 2) patient self-assessment of
treatment efficacy and satisfaction with appearance (Figure 2).
Patient Self-Assessment
The parameters assessed by study subjects in this analysis were the following
questions: a) size of bald spot; b) appearance of hair; c) growth of hair; d) rate of hair
loss; and e) satisfaction with appearance of hair (Table I).
11
Table I. Subject Self Assessment Questions and Rating of Subject Current
Satisfaction.
Self Assessment Questions
1. Since the start of the study, I can see my bald spot getting smaller
. Choice of
answers: strongly agree, agree, no opinion either way, disagree, strongly disagree.
2. Because of the treatment I have received since the start of the study, the
appearance
of my hair is: Choice of answers: a lot better, somewhat better, a little
better, same, a little worse, somewhat worse, a lot worse.
3. Since the start of the study, how would you describe the growth
of your hair?
Choice of answers: greatly increased, moderately increased, slightly increased, no
change, slightly decreased, moderately decreased, greatly decreased.
4. Since the start of the study, how effective do you think the treatment has been
in slowing your hair loss
? Choice of answers: very effective, somewhat effective, not
very effective, not effective at all.
5. Compared to the beginning of the study, which statement best describes your
satisfaction with the appearance
of: a) the hairline at the front of your head? B)
the hair on top of your head? C) your hair overall? Choice of answers: very
satisfied, satisfied, neutral, dissatisfied, very dissatisfied.
Subject Current Satisfaction
1. What is your current level of satisfaction
with your hair condition? Choice of
answers: -3 very dissatisfied,
-2 moderately dissatisfied, -1 slightly dissatisfied, +1 slightly satisfied, +2 moderately
satisfied, +3 very satisfied.
12
Investigative Staff Assessment
As recorded in the Study Protocol Case Report Forms, the primary measure
utilized by the investigative staff in determination of formulation efficacy was a seven
point scale starting with (-3) denoting greatly decreased hair density, ranging to (+3)
marking greatly increased hair density. (Table II).
Table II. Investigator Rating Scale for Subject Assessment in the course of this
study.
Assessment Score
greatly decreased -3
moderately decreased -2
slightly decreased -1
no change 0
slightly increased +1
moderately increased +2
greatly increased +3
Toxicity and Adverse Events
Risk of toxicity in this study was considered minimal inasmuch as the LD/50 for
the botanicals under investigation was far greater than the highest dosages likely to be
consumed. Numerous independent BPH studies evaluating similar formulations support
this observation (Carraro et al. 1996; Wilt et al. 1998; Gerber et al. 1998; Wilt et al. 1999;
Wilt et al. 2000a).
Adverse events were recorded in the Study Protocol Case Report Form Manual
per IRB guidelines. THE CAUSALITY OF ADVERSE EVENTS WAS
based on a four
(4) point scale with 1 = great probability that the adverse event was likely to be related
13
to use of the study drug and 4 = great probability that the adverse event was not likely to
be related to use of the study drug.
The adverse events noted during the course of this study included nausea,
constipation, and diarrhea. However, no adverse events occurring within this pilot were
determined as likely to be related to the use of the study drug. The data are presented
in accordance with the CONSORT clinical study reporting guidelines as described in the
Lancet (Moher et al. 2001).
Results
Study Goals
Our primary goal in this initial pilot study was to assess if the study formulation
was efficacious in arresting scalp hair loss as well as improving hair quality. Based on
anecdotal and open label data, we anticipated being able to show a positive trend
demonstrating a response to therapy. The two measurements utilized for this purpose
were:
1) Investigative staff assessment using a standard assessment
format
Twenty six [26] male subjects, ages 23 to 64 were enrolled in the trial between
July, 1999 and October 1999. Of these 26, nineteen [19] completed the trial. Seven [7]
dropped out prior to study completion, with two of these due to a perceived adverse
event (Table III). Five of the seven patients who withdrew did not report an adverse
event, but dropped out for reasons unrelated to the research study. A total of 19
patients were evaluated after study completion. Duration of participation in this study
ranged from 18 to 24.7 weeks.
14
Table III. Perceived Adverse Events Arising in the Course of the Study.
Complaint Placebo (n=9) Treatment (n=10) Resolved during study Relationship to treatment
Skin 0 1 No Not related
GI 0 1 loss of appetite No Possible
1 flatulence Yes Unlikely
1 diarrhea Yes Unlikely
Neurological 1 0 Yes Placebo
GU 1 frequent urination 0 No Placebo
Miscellaneous 1 aware of heart beat 0 No Placebo
1 heightened sensations
In total, there were 8 perceived adverse events reported by 7
subjects in the course of this study. One subject in the treatment group
reported acne which was present at baseline and worsened during the
course of the study. Three subjects in the treatment group reported
gastroinestinal symptoms, including loss of appetite, flatulence and
diarrhea. One subject in the placebo group reported lightheadedness with
a bowel movement. One subject in the placebo group reported frequent
urination, and one subject in the placebo group reported heightened
sensations and awareness of heartbeat. GI, gastrointestinal; GU,
genitourinary.
On the basis of the investigative staff assessment of change in the patient’s scalp
hair growth from baseline (Figure 1), treatment with the active study formula
demonstrated 60% (6/10) subjects rated as ‘improved’ at the final visit as compared to
baseline. In contrast, only 11% (1/9) in the placebo group were rated as ‘improved’.
These findings show a markedly positive response to treatment as denoted by the
proportion of responders. Due to the small sample size, statistical significance and/or
confidence intervals were not endpoint goals of this pilot study (n = 19) nor were they
achieved, however, future large-scale studies will be designed with these definable
endpoints in mind.
15
2) Subject self-assessment of hair condition
The study participant's self-assessment criterion reflected a time-dependent
analysis of any change noted in the thinning areas of the scalp. Subject assessment of
the “appearance of the bald spot” at the final visit compared to baseline were as follows:
In the treatment group [0]--0% deteriorated, vs. the placebo group where [3]--33%
deteriorated (Figure 2).
Discussion
As previously noted, both LSESr and ß-sitosterol represent, among other
indications, first line BPH treatments in Europe and elsewhere. This is meaningful in
light of the previously noted etiological similarities between these two disorders.
However, to the best of our knowledge, prior to the small trial described in this study, no
controlled and blinded clinical research had previously been conducted on a formula,
either oral or topical, combining LSESr and ß-sitosterol, in the treatment of AGA.
The composition under discussion was designed on the basis of the excellent
safety and efficacy previously demonstrated by each ingredient alone. However, the
published studies were performed on each substance tested separately, but never
together. Nonetheless, both LSESr and ß-sitosterol alone have established clinical
efficacy against BPH.
For example, in a meta-analysis of 18 randomized controlled trials comparing
LSESr to finasteride, involving 2,939 men, both magnitude of improvement and
confidence interval, all tend to favor the efficacy of LSESr with less erectile dysfunction
that that associated with finasteride (Millon et al. 1999). Also, in a randomized, double-
blind trial, BPH patients treated with ß-sitosterol showed a significant improvement in
lower tract urinary symptoms as compared with placebo recipients (Wilt et al. 1998).
Furthermore, a 26-week LSESr vs. finasteride trial involved 1,098 men led to a
remarkable finding which highlights an important safety aspect of these two therapies. It
was found that finasteride lowers the level of prostate specific antigen (PSA) by 30% to
50%. This test is routinely used to screen for prostate cancer and monitor prostate
cancer therapy (Gerber et al. 1998). This study showed that although finasteride does
16
not affect the sequelae of prostate cancer, it may complicate the diagnostic process,
since PSA values must be approximately doubled. In the same study, however, LSESr
demonstrated no impact on PSA levels, suggesting the absence of this potentially
dangerous side effect.
Moreover, in contrast to phytotherapy, where negative side effects are virtually
nonexistent, finasteride has been implicated with the following adverse reactions;
decreased libido (1.8%), erectile dysfunction (1.3%), and ejaculation disorder (1.2%)
(Carraro et al. 1996). Resolution did occur in all men who discontinued therapy with
finasteride due to these side effects. Finally, finasteride treatment is contraindicated in
females as a result of its terotogenic potential. In fact, even the handling of crushed
finasteride tablets by pregnant females is discouraged due to the possibility of systemic
absorption, which may result in risk of feminizing birth defect to a male fetus (Wolf and
Kunte, 1999).
In contrast, these warnings and contraindications have never been associated
with the botanically based substances tested in this study, and in fact LSESr has been
safely and widely used in women for the treatment of dysmenorrhea, genital/urinary
problems, and difficulty association with lactation (Lee et al. 2000). In recognition of the
safe and potentially EFFICACIOUS
nature of the substances under examination, this
small proof of principle trial was conducted with the goal of yielding objective data
demonstrating a positive response for the active formula as measured against placebo.
In summary, the results from this study demonstrate a trend in favor of active
therapy over placebo. The Investigative Staff Assessment showed that 60% (6/10)
subjects were rated as ‘improved’ by the investigator at the final visit as compared to
baseline. This compares to 11% (1/9) in the placebo group (Figure 1). Further, the
Subject Assessment analysis of the ‘appearance of the bald spot’ at the final visit
compared to baseline showed that in the treatment group, no subjects deteriorated. In
contrast however, the placebo group reported three subjects as deteriorated. Inasmuch
as maintenance of hair density and hair quality is a goal for many patients, this finding is
in itself noteworthy.
Based upon the small number of subjects (n = 19) our data justifies
the design of a larger scale study of these substances in the treatment of AGA.
17
In conclusion, we have observed objective evidence of EFFICACY using orally
administered botanical therapy in the treatment of AGA, for the first time. The
implementation of larger gender-specific clinical trials, incorporating local delivery
concomitant with oral administration is planned as an appropriate next step. Research is
also underway to identify additional botanical compounds which may offer useful activity
against AGA.
References
Bracher F. Phytotherapy of benign prostatic hyperplasia. Urologe A 1997; 36:10-17.
Carraro JC, Raynaud JP, Koch G, Chisholm GD, Di Silverio F, Teillac P, Da Silva FC,
Cauquil J, Chopin DK, Hamdy FC, Hanus M, Hauri D, Kalinteris A, Marencak J, Perier A,
Perrin P. Comparison of phytotherapy (Permixon®) with finasteride in the treatment of
benign prostate hyperplasia: A randomized international study of 1,098 patients.
Prostate 1996; 29:231-240.
Delos S, Iehle C, Martin PM, Raynaud J. Inhibition of the activity of ‘basic’ alpha-
reductase (Type 1) detected in DU 145 cells and expressed in insect cells. Steroid
Biochem. Molec. Biol. 1994; 48:347-352.
Gerber G. What is saw palmetto used for, and does it interact with any medications?
Health News 2000; 6:10.
Gerber GS, Zagaja GP, Bales GT, Chodak GW, Contreras BA. Saw palmetto (Serenoa
repens) in men with lower urinary tract symptoms: effects on urodynamic parameters
and voiding symptoms. Urology. 1998; 51:1003-7
Giltay EJ, Gooren LJ. Effects of sex steroid deprivation/administration on hair growth
and skin sebum production in transsexual males and females. Clin Endocrinol Metab
2000; 85:2913-2921.
Guthrie RM, Siegel RL. A multicenter, community-based study of doxazosin in the
treatment of concomitant hypertension and symptomatic benign prostatic hyperplasia:
the Hypertension and BPH Intervention. Trial Clin Ther 1999; 21:1732-1748.
Iehle C, Delos S, Guirou O, Tate R, Raynaud JP, Martin PM. Human prostatic steroid 5
alpha-reductase isoforms--a comparative study of selective inhibitors. J Steroid
Biochem Mol Biol 1995; 54:273-279.
Imperato-McGinley J, Shackleton C, Orlic S, Stoner E. C19 and C21 5 beta/5 alpha
metabolite ratios in subjects treated with the 5 alpha-reductase inhibitor: comparison of
male pseudohermaphrodites with inherited 5 alpha-reductase deficiency. J Clin
Endocrinol Metab 1990; 70:777-782.
18
Itami S, Sonoda T, Kurata S, Takayasu S. Mechanism of action of androgen in hair
follicles. J Dermatol Sci 1994; 7:S98-103.
Kaplan SA, Olsson CA. Patient satisfaction with finasteride in the treatment of
symptomatic benign prostatic hyperplasia. Clin Ther 1996; 18:73-83.
Kaufman KD. Finasteride, 1 mg (Propecia), is the optimal dose for the treatment of men
with male pattern hair loss. Arch Dermatol 1999; 135:989-990.
Klepser TB, Klepser ME. Unsafe and potentially safe herbal therapies. Am J Health Syst
Pharm. 1999; 56:125-38
Lee MM, Lin SS, Wrensch MR, Adler SR, Eisenberg D. Alternative therapies used by
women with breast cancer in four ethnic populations. J Natl Cancer Inst 2000; 92:42-47.
Lise ML, da Gama e Silva TS, Ferigolo M, Barros HM. Abuse of anabolic-androgenic
steroids in sports. Rev Assoc Med Bras 1999; 45:364-370.
Mikolajczyk SD, Millar LS, Wang TJ, Rittenhouse HG, Marks LS, Song W, Wheeler TM,
Slawin KM. "BPSA," a specific molecular form of free prostate-specific antigen, is found
predominantly in the transition zone of patients with nodular benign prostatic
hyperplasia. Urology 2000; 55:41-45.
Millon R, Jacqmin D, Muller D, Guillot J, Eber M, Abecassis J. Detection of prostate-
specific antigen- or prostate-specific membrane antigen-positive circulating cells in
prostatic cancer patients: clinical implications. Eur Urol 1999; 36:278-285.
Moher D, Schulz KF, Altman DG, The CONSORT statement: revised recommendations
for improving the quality of reports of parallel-group randomised trials. Lancet 2001; 357:
1191-94
Morissette J, Durocher F, Leblanc JF, Normand T, Labrie F, Simard J. Genetic linkage
mapping of the human steroid 5 alpha-reductase type 2 gene (SRD5A2) close to
D2S352 on chromosome region 2p23-->p22. Cytogenet Cell Genet 1996; 73:304-307.
Mowszowicz I, Melanitou E, Doukani A, Wright F, Kuttenn F, Mauvais-Jarvis P.
Androgen binding capacity and 5 alpha-reductase activity in pubic skin fibroblasts from
hirsute patients. J Clin Endocrinol Metab 1993; 56:1209-1213.
Negri-Cesi P, Colciago A, Poletti A, Motta M. 5alpha-reductase isozymes and
aromatase are differentially expressed and active in the androgen-independent human
prostate cancer cell lines DU145 and PC3. Prostate 1999; 41:224-232.
Rushton DH, Unger WP, Cotterill PC, Kingsley P, James KC. Quantitative assessment
of 2% topical minoxidil in the treatment of male pattern baldness. Clin Exp Dermatol
1989; 14:40-46.
Sawaya M, Price VH. Different levels of 5alpha-reductase type I and II, aromatase, and
androgen receptor in hair follicles of women and men with androgenetic alopecia. J
Invest Dermatol 1997; 109:296-300.
19
Shapiro J, Wiseman M , Lui H. Practical management of hair loss. Can Fam Phys 2000;
46:1469-1477.
Schilcher H. Is there a rational therapy for symptomatic treatment of benign prostatic
hyperplasia with phytogenic drugs? Illustrated with the example of the prostate agent
from Serenoa repens. Wien Med Wochenschr 1999; 149:236-240.
Swoboda H, Kopp B. Serenoa repens--the saw palmetto or dwarf palm. Wien Med
Wochenschr 1999; 149:235.
Wang HX, Ng TB. Natural products with hypoglycemic, hypotensive,
hypocholesterolemic, antiatherosclerotic and antithrombotic activities. Life Sci 1999;
65:2663-2677.
Wilson JD, Roehrborn C. Long-term consequences of castration in men: lessons from
the Skoptzy and the eunuchs of the Chinese and Ottoman courts. J Clin Endocrinol
Metab 1999; 84:4324-4331.
Wilt T, Ishani A, Stark G, MacDonald R, Mulrow C, Lau J. Serenoa repens for benign
prostatic hyperplasia. Cochrane Database Syst Rev 2000a; 2:CD001423.
Wilt T, Mac Donald R, Ishani A, Rutks I, Stark G. Cernilton for benign prostatic
hyperplasia. Cochrane Database Syst Rev 2000b; 2:CD001042.
Wilt TJ, MacDonald R, Ishani A. Beta-sitosterol for the treatment of benign prostatic
hyperplasia. BJU Int 1999; 83: 976-983.
Wilt TJ, Ishani A, Stark G, MacDonald R, Lau J, Mulrow C. Saw palmetto extracts for
treatment of benign prostatic hyperplasia: a systematic review. JAMA. 1998; 280:1604-9
Wolff H, Kunte C. Therapy of androgenetic alopecia with finasteride. What must be
considered in consultation and drug prescribing. MMW Fortschr Med 1999; 141:38-40.
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Figure Legends
Figure 1. Graphic representation of the results of the Investigative Staff Assessment of
the study group. The blue (upper) bar represents subjects treated with active
formulation, versus the green (lower) bar representing subjects in the placebo group.
The X-axis represents numbers of subjects
(1-10). The results demonstrated that 6 of 10 subjects (60%) were rated as improved in
the group taking the active formulation, compared to 1 of 9 subjects (11%) in the placebo
control group.
Fig. 1
Figure 2. Graphic representation of the results of the Subjects’ Self-Assessment of the
study group. The criterion represented here was the perceived deterioration of the bald
spot. The yellow (left-hand) bar represents subjects in the treatment group, versus the
blue (right hand) bar representing subjects in the placebo group. The Y-axis represents
percent deterioration. The results demonstrated that 0 of 10 subjects (0%) in the group
taking active formulation reported any deterioration of the bald spot, compared to 3 of 9
subjects (33%) in the placebo control group.
Fig. 2
... In a randomized, double-blind, placebo-controlled trial, a softgel containing 50 mg β-sitosterol and 200 mg saw palmetto extract or placebo was administered twice daily in 19 male AGA patients, aged between 23 and 64 years. 8 After 21 weeks, the blinded investigators reported that 60% of patients had "improved outcome" in the treatment arm compared to 11% receiving placebo. 8 The blinded investigator rating scale ranged from greatly decreased to greatly increased hair, from À3 to +3. ...
... 8 After 21 weeks, the blinded investigators reported that 60% of patients had "improved outcome" in the treatment arm compared to 11% receiving placebo. 8 The blinded investigator rating scale ranged from greatly decreased to greatly increased hair, from À3 to +3. However, the trial lacked an appropriate definition of "improved outcome" and statistical significance. ...
... However, the trial lacked an appropriate definition of "improved outcome" and statistical significance. 8 Three of 10 patients in the treatment arm experienced gastrointestinal side effects (loss of appetite, flatulence, and diarrhea; Table 1). 8 An open-label study compared the effectiveness of Serenoa repens (saw palmetto) with finasteride in treating mild to moderate male AGA in 100 patients (age range: 20-40 years; Table 1). ...
Article
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Androgenetic alopecia (AGA) is the most common cause of hair loss in men, often requiring medical attention. The US FDA approved topical minoxidil and oral finasteride to treat AGA. Topical minoxidil requires a long-term application to observe improvement; oral finasteride may cause undesirable side effects. Therefore, natural products may be an alternative when patients are skeptical about these two conventional treatments. Physicians may also suggest natural products in conjunction with topical minoxidil or oral finasteride to enhance clinical outcomes. This article reviews the prospect of natural products in treating male AGA. A systematic search was conducted in PubMed, CINAHL, Scopus, Web of Science, and EMBASE (Ovid) on July 19, 2021. In addition, the bibliographies of selected articles were hand-searched to identify relevant studies. After deduplication and screening, 11 clinical studies meet the criteria for detailed review. The selected clinical studies suggest that saw palmetto, caffeine, melatonin, marine extracts, rosemary oil, procyanidin, pumpkin seed oil, and cannabidiol (CBD) oil might be considered in male AGA treatment.
... All phytosterols have anti-androgenic effects and decrease tissue sensitivity to androgens, besides, androgen activity is decreased by inhibiting 5-alpha reductase Inhibition of this enzyme reduces the conversion of testosterone to dihydrotestosterone, an active form of this hormone in tissues (18) . In addition, phytosterols can treat benign prostatic hyperplasia (BPH) in the prostate by reducing testosterone and dihydrotestosterone levels. ...
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Purpose: Oilseeds and their related products are known to have various bioactive and health-promoting ingredients. In this research, we investigated the effects of phytosterols and fatty acids of Pistacia vera on spermatogenesis process and testis histological changes in Wistar male rats for the first time. Materials and methods: A total number of 64 adult male Wistar rats were divided randomly into eight groups including one control group, and seven test groups. Test groups received phytosterols, fatty acids, and pistachio oil orally for 30 days. Then, LH, FSH and serum testosterone levels were determined. Also, the spermatogenesis process and changes in testicular tissue in rats were investigated. Results: The results of this research suggest that phytosterols in doses 10 and 50 mg/kg reduce spermatogenesis process. Fatty acid in a low dose of 10 mg/kg increases spermatogenesis, but when a high dose of 50 mg/kg was used, it harmed the spermatogenesis process. When low levels of phytosterols and fatty acids are used simultaneously in dose 5 mg/kg, improvement in spermatogenesis process is observed but when these were used together in the dose of 25 mg/kg, the spermatogenesis process was disrupted. Using pistachio oil alone also improved spermatogenesis process. Conclusion: It seems that phytosterols reduce spermatogenesis at high and low doses, while fatty acids increase spermatogenesis when used in low doses and reduce this process when used in high doses. The use of fatty acids extracted from pistachios to treat infertility in men seems hopeful.
... [140][141][142] In a study with 10 male subjects with AGA, improvement was noted in 60% of the participants. 143 Another study applying topical SR extract in lotion and shampoo for 3 months led to 35% increase in hair density. 140 Rossi et al. conducted an open-label study enrolling 100 male patients to study the efficacy of Serenoa repens 320 mg daily vs. finasteride 1 mg daily for 24 months. ...
Article
Full-text available
Background Androgenetic alopecia (AGA) is the most common form of hair loss consisting of a characteristic receding frontal hairline in men and diffuse hair thinning in women, with frontal hairline retention, and can impact an individual's quality of life. The condition is primarily mediated by 5-alpha-reductase and dihydrotestosterone (DHT) which causes hair follicles to undergo miniaturization and shortening of successive anagen cycles. Although a variety of medical, surgical, light-based and nutraceutical treatment options are available to slow or reverse the progression of AGA, it can be challenging to select appropriate therapies for this chronic condition. Aims To highlight treatment options for androgenetic alopecia taking into consideration the efficacy, side effect profiles, practicality of treatment (compliance), and costs to help clinicians offer ethically appropriate treatment regimens to their patients. Materials and Methods A literature search was conducted using electronic databases (Medline, PubMed, Embase, CINAHL, EBSCO) and textbooks, in addition to the authors' and other practitioners' clinical experiences in treating androgenetic alopecia, and the findings are presented here. Results Although topical minoxidil, oral finasteride, and low-level light therapy are the only FDA-approved therapies to treat AGA, they are just a fraction of the treatment options available, including other oral and topical modalities, hormonal therapies, nutraceuticals, PRP and exosome treatments, and hair transplantation. Discussion Androgenetic alopecia therapy remains challenging as treatment selection involves ethical, evidence-based decision-making and consideration of each individual patient's needs, compliance, budget, extent of hair loss, and aesthetic goals, independent of potential financial benefits to the practitioners.
... The ability of β-sitosterol and other phytosterols to inhibit aromatase and 5-alphareductase has been well documented, and this inhibitory capacity has been exploited to treat pathologies such as benign prostatic hyperplasia and androgenetic alopecia [26][27][28]. One of the richest sources of phytosterols, particularly β-sitosterol, is Serenoa repens, whose extracts have been largely studied and used in nutraceutical formulations (Permixon, Calprost, Difaprost) proposed for the adjunct therapy of benign prostatic hyperplasia [25,29,30]. ...
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Cannabis sativa L. has been used for a long time to obtain food, fiber, and as a medicinal and psychoactive plant. Today, the nutraceutical potential of C. sativa is being increasingly reappraised; however, C. sativa roots remain poorly studied, despite citations in the scientific literature. In this direction, we identified and quantified the presence of valuable bioactives (namely, β-sitosterol, stigmasterol, campesterol, friedelin, and epi-friedelanol) in the root extracts of C. sativa, a finding which might pave the way to the exploitation of the therapeutic potential of all parts of the C. sativa plant. To facilitate root harvesting and processing, aeroponic (AP) and aeroponic-elicited cultures (AEP) were established and compared to soil-cultivated plants (SP). Interestingly, considerably increased plant growth—particularly of the roots—and a significant increase (up to 20-fold in the case of β-sitosterol) in the total content of the aforementioned roots’ bioactive molecules were observed in AP and AEP. In conclusion, aeroponics, an easy, standardized, contaminant-free cultivation technique, facilitates the harvesting/processing of roots along with a greater production of their secondary bioactive metabolites, which could be utilized in the formulation of health-promoting and health-care products.
... Finasteride is an FDA approved type II 5-α reductase inhibitor for treating alopecia in men and benign prostatic hyperplasia (BPH) Fig 3. Dutasteride, Global Pharmaceutical Sciences Review (GPSR) alfatradiol are also used for alopecia management (Prager, Bickett, French, & Marcovici, 2002). ...
Article
Alopecia is an autoimmune and non-communicable disorder. The immune system, especially Tlymphocytes, present around the hair follicle attacks, cause inflammation and loss of hair from some or all areas of the scalp. It is highly prevalent in the United States; an estimated 4.5 million are affected by this disease. Finasteride and minoxidil are mostly used to treat alopecia in the form of solutions and gels. However, their use has been limited due to severe adverse effects observed during therapy. This article summarizes the epidemiology, risk factors, pathophysiology, sign and symptoms, and diagnostic parameters with a focus on novel treatment strategies.
... The ability of β-sitosterol and other phytosterols to inhibit aromatase and 5-alphareductase has been well documented, and this inhibitory capacity has been exploited to treat pathologies such as benign prostatic hyperplasia and androgenetic alopecia [26][27][28]. One of the richest sources of phytosterols, particularly β-sitosterol, is Serenoa repens, whose extracts have been largely studied and used in nutraceutical formulations (Permixon, Calprost, Difaprost) proposed for the adjunct therapy of benign prostatic hyperplasia [25,29,30]. ...
Preprint
Cannabis Sativa L. has been used for a long time to obtain food, fiber and as a medicinal and psychoactive plant. Today the nutraceutical potential of C. Sativa is being increasingly reappraised; however, C. Sativa roots remain poorly studied, despite citations in the scientific literature. In this direction, we identified and quantified the presence of valuable bioactives (namely β-sitosterol, stigmasterol, campesterol, friedelin and epi-friedelanol) in the root extracts of C. Sativa, a finding which might pave the way to the exploitation of the therapeutic potential of C. Sativa in all its parts. To facilitate roots harvesting and processing, aeroponic (AP) and aeroponic elicited cultures (AEP), have been set up and compared to soil-cultivated plant (SP): interestingly a considerable overgrowth of the plants - particularly of roots - and a significant increase (up to 20 fold in the case of β-sitosterol) in the total content of the above roots’ bioactive molecules have been observed in AP and AEP. In conclusion aeroponics, an easy, standardised, free of contaminant cultivation tecchnique, allows an ease harvesting/processing of roots along with a greater production of their secondary bioactive metabolites which could be utilized in the formulation of health promoting and health care products.
Article
Full-text available
Cannabis sativa var. Kompolti, a variety routinely used for food production purposes, is characterized by a low concentration of psychoactive molecules, although containing many other biologically attractive metabolites in all parts of the plant, including the roots. In the present work, we evaluate the specific biological activities of the roots’ extract from plants cultivated through aeroponics, an affordable and reliable method facilitating the isolation and processing of roots, with the advantage of being suitable for industrial scale-up. Furthermore, aeroponics results in an increased net accumulation of the most biologically attractive constituents (β-sitosterol, friedelin and epi-friedelanol) found in the roots. The ethanolic extract of the aeroponic roots of C. sativa (APEX) and its separate components are studied to evaluate their anti-inflammatory (modulation of the expression level of specific markers upon LPS stimulation in U937 cells, such as IL-6, IL-8, TNF-α, IkB-α, iNOS, IRAK-1 and miR-146a) and antioxidant (in either acellular or cellular settings) activities. The APEX anti-inflammatory and antioxidant capacities are also functionally benchmarked using the wound-healing assay. On the whole, the data obtained show that APEX and its main components showed significant anti-inflammatory and antioxidant activities, which may render the exploitation of roots as a source of natural antioxidants and anti-inflammatory agents highly attractive, with the additional technical and economic advantages of aeroponics compared to soil cultivation.
Article
Full-text available
Background: Dihydrotestosterone (DHT), the most critical pathogenic androgen in hair loss, is identified as an etiologic factor of androgenetic alopecia (AGA). The AGA is a genetically common disorder among men and is characterized by the progressive conversion of hair follicles into small vellus hair. Steroid 5 alpha-reductase type 1 (5AR1) is a crucial target responsible for this gradual replacement. The 5AR1 function is determined by converting testosterone to DHT. The inhibitors of 5AR1 play their role by blocking the DHT production pathway. Objectives: This study focused on the potent inhibitors of the 5AR1 enzyme to suggest effective synthetic drugs for restoring hair loss with fewer side effects. Methods: The three-dimensional structure of 5AR1 was created using homology modeling methods. Then, the inhibitory effects of some significant compounds from natural sources were examined on the 5AR1 protein using molecular docking approaches. Results: The obtained results suggest that two natural compounds isolated from Serenoa repens, including beta-sitosterol and stigmasterol, could inhibit the regular activity of 5AR1 and can be recommended as safe and novel AGA medicines for hair restoration.
Article
Background: Dihydrotestosterone (DHT), the most critical pathogenic androgen in hair loss, is identified as an etiologic factor of androgenetic alopecia (AGA). The AGA is a genetically common disorder among men and is characterized by the progressive conversion of hair follicles into small vellus hair. Steroid 5 alpha-reductase type 1 (5AR1) is a crucial target responsible for this gradual replacement. The 5AR1 function is determined by converting testosterone to DHT. The inhibitors of 5AR1 play their role by blocking the DHT production pathway. Objectives: This study focused on the potent inhibitors of the 5AR1 enzyme to suggest effective synthetic drugs for restoring hair loss with fewer side effects. Methods: The three-dimensional structure of 5AR1 was created using homology modeling methods. Then, the inhibitory effects of some significant compounds from natural sources were examined on the 5AR1 protein using molecular docking approaches. Results: The obtained results suggest that two natural compounds isolated from Serenoa repens, including beta-sitosterol and stigmasterol, could inhibit the regular activity of 5AR1 and can be recommended as safe and novel AGA medicines for hair restoration.
Article
Objectives: The hair follicle is composed of more than 20 kinds of cells, and mesoderm derived dermal papilla cells and keratinocytes cooperatively contribute hair growth via Wnt/β-catenin signaling pathway. We are to investigate β-catenin expression and regulatory mechanism by CBD in alopecia hair tissues and dermal papilla cells. Methods: We performed structural and anatomical analyses on alopecia patients derived hair tissues using microscopes. Pharmacological effect of CBD was evaluated by β-catenin expression using RT-PCR and immunostaining experiment. Results: Morphological deformation and loss of cell numbers in hair shaft were observed in alopecia hair tissues. IHC experiment showed that loss of β-catenin expression was shown in inner shaft of the alopecia hair tissues, indicating that β-catenin expression is a key regulatory function during alopecia progression. Consistently, β-catenin expression was decreased in testosterone or PMA treated dermal papilla cells, suggesting that those treatments are referred as a model on molecular mechanism of alopecia using dermal papilla cells. RT-PCR and immunostaining experiments showed that β-catenin expression was decreased in RNA level, as well as decreased β-catenin protein might be resulted from ubiquitination. However, CBD treatment has no changes in gene expression including β-catenin, but the decreased β-catenin expression by testosterone or PMA was restored by CBD pretreatment, suggesting that potential regulatory effect on alopecia induction of testosterone and PMA. Conclusion: CBD might have a modulating function on alopecia caused by hormonal or excess of signaling pathway, and be a promising application for on alopecia treatment.
Article
Two steroid 5α-reductase isoenzymes catalyze the conversion of testosterone into dihydrotestosterone, the more bioactive androgen, which is essential for male phenotypic sexual differentiation and for androgen-mediated growth of such tissues and organs as the prostate. Inherited mutations in SRD5A2 cause male pseudohermaphroditism. The SRD5A1 and SRD5A2 genes encoding the steroid 5α-reductase type 1 and type 2 isoenzymes have been previously assigned by in situ hybridization to 5p15 and 2p23, respectively. To map the SRD5A2 gene by linkage analysis, a novel RsaI RFLP detected in exon I and a TA repeat polymorphism found in exon V were genotyped in eight CEPH reference families. A two-point linkage analysis was performed between these polymorphisms and the chromosome 2 microsatellite markers of Généthon and NIH/CEPH. The closest linkage was observed with D2S352 (Zmax = 24.06; θmax = 0.001) in the region 2p23→p22. To further define the localization of SRD5A2, a framework map, including nine Généthon markers flanking the polymorphic SRD5A2 locus, was built by multipoint linkage analysis. This led to a high-resolution genetic map of the region flanking the polymorphic SRD5A2 gene, including the nine Généthon markers and three NIH/CEPH markers, yielded the following order: tel-D2S48-D2S149-D2S320-D2S171-D2S165-[D2S352/SRD5A2]-D2S367-[D2S19/D2S177]-[D2S391/ CALM]-D2S378-cen.Copyright © 1996 S. Karger AG, Basel
Article
Objectives To conduct a systematic review of the evidence for the efficacy of β-sitosterolin men with symptomatic benign prostatic hyperplasia (BPH).Methods Studies were identified through Medline™ (1966–98), EMBASE™ , Phytodok, the Cochrane Library, bibliographies of identified trials and review articles, and contact with study authors and pharmaceutical companies. Randomized trials were included if: men had symptomatic BPH; plant extract preparations contained β-sitosterols; a control group received placebo or a pharmacological therapy; and treatment duration was ≥30 days. Study characteristics, demographic information, enrolment criteria and outcomes were extracted.ResultsFour trials comprising a total of 519 men met the inclusion criteria. All were double-blind and lasted 4–26 weeks. Three studies used nonglucosidic β-sitosterols and one used a preparation that contained only β-sitosterol-β-d-glucoside. Compared with placebo, β-sitosterolimproved urinary symptom scores and flow measures. For the two studies reporting the International Prostate Symptom Score (IPSS), the weighted mean difference (WMD) against placebo was −4.9 IPSS points (95% confidence interval, CI,−6.3 to−3.5). The WMD for peak urinary flow rate was 3.91 mL/s (95% CI 0.91 to 6.90, four studies) and for residual volume the WMD was −28.62 mL (95% CI−41.42 to−15.83, four studies). β-sitosteroldid not reduce prostate size. The trial using pure β-sitosterol-β-d-glucoside (WA184) showed no improvement in urinary flow measures. Withdrawal rates for men assigned to β-sitosterol and placebo were 7.8% and 8.0% (not significant), respectively.Conclusionβ-sitosterolimproves urological symptoms and flow measures. However, the existing studies are limited by short treatment duration and lack of standardized β-sitosterol preparations. Their long-term effectiveness, safety and ability to prevent the complications of BPH are unknown.
Article
Phytopharmaceutical agents have been used for a long time in the treatment of symptomatic benign prostatic hyperplasia (BPH). However, until recently, it has been questioned whether phytotherapy is superior to a placebo treatment. In this article, the most widely used phytopharmaceutical agents, such as saw palmetto berry extracts, Radix urticae extracts, pumpkin seeds, pollen extracts and different phytosterols, are described. In addition, both in vitro and in vivo studies are discussed in an attempt to explain a possible mechanism of action. There are several new clinical studies which demonstrate a significant benefit compared with placebo treatment. Based on these results, the use of phytopharmaceutical agents for the treatment of mild to moderate symptomatic BPH seems to be well justified. So far, no significant inhibition of further prostate growth has been demonstrated. For this, a careful follow-up of the patients is necessary so as not to miss a deterioration and perhaps the need for an operation.
Article
Male subjects administered the 5 alpha-reductase inhibitor finasteride were studied to determine its effect on C19 and C21 5 alpha-metabolism. Plasma testosterone (T) and 5 alpha-dihydrotestosterone (DHT) were measured and T/DHT ratios determined at doses of 0.2-80 mg. Urinary etiocholanolone (5 beta)/androsterone (5 alpha) ratios and 5 beta/5 alpha metabolite ratios of cortisol, 11 beta-hydroxyandrostenedione, and corticosterone were also measured. The steroid profile was compared to male pseudohermaphrodites with inherited 5 alpha-reductase deficiency who have a global defect in C19 and C21 5 alpha-metabolism. The mean plasma DHT levels were decreased at all doses, resulting in elevated T/DHT ratios. The mean urinary etiocholanolone/androsterone, 11 beta-hydroxyetiocholanolone/11 beta-hydroxyandrosterone, tetrahydrocortisol/allotetrahydrocortisol, and tetrahydrocorticosterone/allotetrahydrocorticosterone ratios were elevated compared to pretreatment levels and placebo control values. The mean ratios appeared to be dose dependent for plasma T/DHT, urinary etiocholanolone/androsterone tetrahydrocorticosterone/allotetrahydrocorticosterone ratios. The mean 11 beta-hydroxyetiocholanolone-hydroxyandrosterone ratio was maximally elevated at the lowest doses. The results indicate that finasteride has a broad steroid spectrum inhibiting C19 and C21 5 alpha-steroid metabolism and affecting hepatic and peripheral 5 alpha-metabolism. These results suggest that a single gene codes for a single 5 alpha-reductase enzyme with affinity for multiple steroid substrates. The steroid profile is strikingly similar to that of male pseudohermaphrodites with inherited 5 alpha-reductase deficiency.
Article
Forty-seven men with male pattern baldness were treated in a double-blind clinical trial with topical 2% minoxidil or placebo. Twelve were randomly selected for quantitative hair measurement using the unit area trichogram and visual counting. There was no significant difference after 6 or 12 months of treatment with a 2% minoxidil solution for total hair density (THD; hair cm-2), meaningful hair density (MHD; hair greater than 40 microns in diameter greater than 30 mm in length cm-2), per cent of hair in the anagen growth phase, or the per cent of meaningful hair in the anagen growth phase. Significantly fewer hairs were recorded with the visual hair counting method, compared to values obtained from adjacent sites with the unit area trichogram. In addition, a significantly larger mean total hair count was recorded by an experienced observer, compared to an inexperienced observer. Increased pigmentation was observed within the vellus hair population of treated subjects. Our findings indicate that minoxidil appears unlikely to affect the long-term course of male pattern baldness. However, we found no significant deterioration in total hair density, or meaningful hair density in treated subjects, suggesting minoxidil may have a prophylactic effect. Further long-term studies employing the unit area trichogram are required to evaluate this finding.
Article
We have measured the total (cytosolic plus nuclear) androgen binding capacity of pubic skin fibroblasts from nine patients with hirsutism of various origin. Confluent intact cell monolayers were incubated with increasing concentrations (0.05-2 nM) of [3H]dihydrotestosterone ([3H]DHT) with or without a 200-fold excess of unlabeled DHT. The androgen binding capacities (mean +/- SD) were similar in normal men (411 +/- 171 fmol/mg DNA), women (310 +/- 103 fmol/mg DNA), and hirsute patients (313 +/- 141 fmol/mg DNA) regardless of the plasma androgen levels. In contrast, the 5 alpha-reductase level in pubic skin fibroblasts (mean +/- SD) was, as previously described, higher in hirsute women (3.3 +/- 2.6 fmol/micrograms DNA . h) than in normal women (1.1 +/- 0.6 fmol/microgram DNA . h; P less than 0.05). We conclude from these data that: 1) increased androgen binding capacity cannot be held responsible for hypersensitivity to androgens in hirsutism; 2) the androgen receptor is not regulated by androgens in human skin, as similar levels are observed in men, women, and hirsute patients; 3) this contrasts with 5 alpha-reductase activity and emphasizes the importance of this enzyme as an amplifier of androgen action in areas where it is stimulated by androgens, such as pubic skin.
Article
The present study describes the independent expression of the type 1 and 2 isoforms of human 5 alpha-reductase in the baculovirus-directed insect cell expression system and the selectivity of their inhibition. The catalytic properties and kinetic parameters of the recombinant isozymes were consistent with published data. The type 1 isoform displayed a neutral (range 6-8) pH optimum and the type 2 isoform an acidic (5-6) pH optimum. The type 2 isoform had higher affinity for testosterone than did the type 1 isoform (Km = 0.5 and 2.9 microM, respectively). Finasteride and turosteride were selective inhibitors of the type 2 isoform (Ki (type 2) = 7.3 and 21.7 nM compared to Ki (type 1) = 108 and 330 nM, respectively). 4-MA and the lipido-sterol extract of Serenoa repens (LSESr) markedly inhibited both isozymes (Ki (type 1) = 8.4 nM and 7.2 micrograms/ml, respectively; Ki (type 2) = 7.4 nM and 4.9 micrograms/ml, respectively). The three azasteroids were competitive inhibitors vs substrate, whereas LSESr displayed non-competitive inhibition of the type 1 isozyme and uncompetitive inhibition of the type 2 isozyme. These observations suggest that the lipid component of LSESr might be responsible for its inhibitory effect by modulating the membrane environment of 5 alpha-reductase. Partially purified recombinant 5 alpha-reductase type 1 activity was preserved by the presence of lipids indicating that lipids can exert either stimulatory or inhibitory effects on human 5 alpha-reductase.
Article
In order to investigate the mode of action of testosterone (T) on human hair follicles we studied the metabolism of T and localization of androgen receptors in outer root sheath cells (ORSC) and dermal papilla cells (DPC) from different body sites. T was principally metabolized to androstenedione (delta 4) even in beard ORSC as well as epidermal keratinocytes (EK), and the ratio of apparent 5 alpha-reductase (5 alpha-R) to 17 beta-hydroxysteroid dehydrogenase (17 beta-HSD) did not differ between these two kinds of cells. The 5 alpha-R activity in beard DPC was 3 times as high as that in occipital scalp and axillary DPC. The 5 alpha-R of beard DPC exhibited a narrow optimum pH of 5.5, which is characteristic of type 2 enzyme present in androgen target cells. In contrast, 5 alpha-R of DPC from axillary and occipital scalp hair showed a broad optimum pH range between 6.5-9.0 corresponding to type 1 5 alpha-R. Androgen receptors were detected in the DPC of beard and axillary hair follicles, but not in those of occipital scalp hair follicles using immunohistochemical staining with polyclonal anti-androgen receptor antibody. Epithelial cells of the hair bulb were not stained by the antibody. Androgen receptors were also detected in the nuclei of cultured beard and axillary DPC, but the DPC from occipital scalp hair follicles showed little staining with the antibody. We also examined the effects of T on the DNA synthesis and proliferation of cultured ORSC and DPC. T did not have a proliferative effect on either type of cell when cultured alone.(ABSTRACT TRUNCATED AT 250 WORDS)
Article
The purpose of this study was 2-fold: (1) to identify the 5 alpha-reductase (5 alpha-R) isozyme(s) present in DU 145 cells, a human cell-line of low androgen sensitivity derived from a cerebral metastasis of an epithelial prostate cancer; and (2) to compare the inhibitory potencies of three compounds on the 'basic' 5 alpha-R isozyme expressed in a baculovirus-directed insect cell system. Conversion of testosterone (T) into 5 alpha-dihydrotestosterone (DHT) in DU 145 cells was measured by HPLC coupled to a Flo-one HP radioactivity detector. DU 145 cells exhibited 5 alpha-R activity (21 pmol DHT/min/mg protein) at pH 7.4 which disappeared at pH 5.5 suggesting that, of the two genomically distinct human isozymes identified so far, type 1 5 alpha-R is expressed in DU 145 cells. This was confirmed by at least two observations: first, 5 alpha-R activity in DU 145 cells was inhibited with much higher potency by 4-MA than by finasteride which is known to be a very poor competitor of the 'basic' enzyme (IC50s = 2.8 +/- 0.2 and 264 +/- 55 nM, respectively). Second, only the type 1 5 alpha-R cDNA and not type 2 5 alpha-R cDNA hybridized with DU 145 RNA. A high potency differential was also recorded for the inhibition of 'basic' type 1 5 alpha-R expressed in a baculovirus-directed-insect cell system by these two compounds, 4-MA being considerably more active than finasteride (Ki = 8.4 +/- 2.3 and 330 +/- 9 nM, respectively). This inhibition was competitive. On the other hand, inhibition by an n-hexane lipid/sterol extract of Serenoa repens (LSESr) was non-competitive and, when expressed in terms of recommended therapeutic doses, was 3-fold greater for LSESr than for finasteride. These studies suggest that LSESr might exert a regulatory inhibitory activity due to its specific lipid/sterol composition.