A retrospective study of survival in breast cancer patients undergoing deuterium depletion in addition to conventional therapies

Abstract

There is increasing evidence that the heavy isotope of hydrogen, deuterium (D), has a pivotal role in cell signalling and that its depletion through the replacement of normal drinking water with deuterium-depleted water (DDW) results in tumour necrosis. The impact of D–depletion on breast cancer outcome was studied retrospectively. The normal daily water intake (150 ppm D) of 232 breast cancer patients was replaced with DDW (65-105 ppm D) for at least 91 days, without altering conventional treatment regimens. According to staging at initial diagnosis, patients with early stage breast cancer (n158) achieved a median survival time (MST) of 217 months (18.1 years), compared with 52 months (4.3 years) in patients with advanced disease (n74). The MST is pending in the subgroup of patients who were in remission at the start of DDW treatment; only one out of 48 patients died during the cumulative follow-up period of 221.1 years. Although single DDW treatment was effective, an outstandingly long MST of 24.4 years was attained in the subgroup of 53 patients who were treated with DDW at least twice. In comparison with published data, DDW treatment in combination with or as an extension of conventional therapies noticeably prolonged MST in certain subgroups of breast cancer patients. D-depletion may also be a highly effective therapy for preventing the recurrence of breast cancer. Furthermore, the method is safe and can be easily integrated into standard treatment regimens for breast cancer.

Full text

Introduction
Breast cancer is a major health problem worldwide and
the second most common cause of cancer-related deaths in
women. Although population screening by mammography
can ensure early detection of breast cancer and the
rapid development of therapeutic options (i.e. surgery,
radiotherapy, hormonal therapy, chemotherapy, more
recently, targeted cancer drugs) provides the possibility
of multidisciplinary treatment, the morbidity of breast
cancer is still high [1]. Trends in mortality and morbidity
did not show a decline in the United States between
2005 and 2009 [2]. Similarly, age-standardised incidence
rates for breast cancer in females in Great Britain did not
decrease in the past 7 to 8 years [3]. Long-term follow-
up studies showed that the probability of relapse ranges
from 30 to 85% depending on tumour stage [4]. Median


chemotherapy and targeted biological therapies are
the mainstay of treatment for metastatic breast cancer
Journal of Cancer
Research & Therapy
Original research

9
A retrospective study of survival in breast cancer patients
undergoing deuterium depletion in addition to conventional
therapies
Krempels K1, Somlyai I1, Gyöngyi Z2, Ember I2, Balog K1, Abonyi O1 and Somlyai G1,
1 
2 Department of Public Health, Medical School, University of Pécs, Pécs, 7624, Hungary
Abstract
There is increasing evidence that the heavy isotope of hydrogen, deuterium (D), has a pivotal role in cell signalling and that its depletion
through the replacement of normal drinking water with deuterium-depleted water (DDW) results in tumour necrosis. The impact of

patients was replaced with DDW (65-105 ppm D) for at least 91 days, without altering conventional treatment regimens. According
to staging at initial diagnosis, patients with early stage breast cancer (n158) achieved a median survival time (MST) of 217 months
(18.1 years), compared with 52 months (4.3 years) in patients with advanced disease (n74). The MST is pending in the subgroup of
patients who were in remission at the start of DDW treatment; only one out of 48 patients died during the cumulative follow-up period

of 53 patients who were treated with DDW at least twice. In comparison with published data, DDW treatment in combination with or


integrated into standard treatment regimens for breast cancer.
Keywords: deuterium depletion; deuterium-depleted water; breast cancer; retrospective study; median survival time; early stage
breast cancer; advanced disease
Corresponding author:
      36-1-381-
 gsomlyai@hyd.
hu
Received 26 May 2013 Revised 6 September 2013 Accepted 16 September
2013 Published 25 September 2013
Citation: Krempels K, Somlyai I, Gyöngyi Z, Ember I, Balog K, Abonyi
O, Somlyai G (2013) A retrospective study of survival in breast cancer
patients undergoing deuterium depletion in addition to conventional

29.
Copyright: 2013 Krempels K, et al. This is an open-access article
        
License, which permits unrestricted use, distribution and reproduction
in any medium, provided the original author and source are credited.
and the goal in such cases is to maintain quality of life,
ameliorate symptoms, and delay progression [5].
A wide range of information is available regarding the
     
(D) and protium (H), the two stable isotopes of hydrogen
Open Access
NobleResearch
www.nobleresearch.org

195 
[6, 7, 8]. The possible role of naturally occurring
      
       
biological systems [9, 10, 11]. The in vitro growth rate of
      
in culture media prepared with deuterium-depleted
water (DDW), and the administration of DDW as drinking
water resulted in complete or partial tumour regression
      


was demonstrated both in vitro [10] and in vivo [11].



   
given DDW to drink [12]. The registered oral formulation
of the deuterium-depleted veterinary anticancer drug
Vetera-DDW-25 AUV has been used successfully in
the treatment of household pets, mainly dogs and
cats with spontaneous malignancies. Siniak et al. also


 

in a double-blind, randomised, 4-month-long, human
      
follow-up and retrospective evaluation of 91 prostate
cancer patients suggested that the method might reduce
the mortality rate of prostate cancer because it delayed
disease progression and prolonged MST [14]. Lung
cancer patients undergoing DDW treatment in addition to
conventional treatments achieved longer MSTs, and DDW
was also successfully applied in lung cancer complicated
by brain metastases [15].
The aim of the present retrospective study was to
investigate the impact of D-depletion on the outcome of
breast cancer. The daily water intake of 232 breast cancer
patients was replaced with DDW (105-65 ppm D) for at
least 91 days, without altering the conventional treatment
regimens. The primary end point of the study was MST. To
obtain data comparable to historical controls, subgroups
were created based on initial staging of the disease.
Materials and methods
DDW production
DDW was produced from ordinary water containing
a natural amount of D (150 ppm), equivalent to (16.8
      
D-concentration to 105-65 ppm. The production of DDW
2O)
and heavy water (D2O). At the boiling point of normal water
the steam in equilibrium with the liquid phase contains
     
evaporation steps, which are carried out in distillation
towers for industrial quantities, the deuterium content
of water can be decreased commensurate with the tray
number of the distillation tower. D-concentration was

1 ppm precision [16].
Main principles of administration of DDW
Per os (PO) DDW treatment supplemented and did not
replace conventional therapy. The aim of the treatment
       
by replacing the normal water intake with DDW as
described previously [14]. DDW treatment started at 105
ppm D, and was gradually decreased to 85 ppm and 65
ppm (with each step 20 ppm lower than the previously
applied preparation) every 1 to 3 months, for a total
treatment period of 6 to 10 months. DDW treatment
was discontinued for 2 to 3 months and started again for
repeated 4- to 6-month treatment durations. Over time,
the treatment periods were shortened, although each
had to be at least 3 months long, and the breaks were
      
depletion unit) was calculated according to the following

General characterisation of patients
According to the study protocol, eligible patients
were women who were diagnosed with histologically
  
for at least 91 days. The study included 232 patients who
met the inclusion criteria (Table 1). The median age of
the study population was 50 years. The patients started
       
median length of time between diagnosis and start of
DDW treatment (DG_to_DDWstart) was 13.2 months. The
standard deviation (SD) of the average length of time shows
that in some cases several years elapsed until inclusion in
the study whereas other patients were enrolled after only
a few months. The median duration of DDW treatment
was 14.1 months (LoDDW) and the cumulative duration
of DDW treatment was 474 years. The cumulative length
of total follow-up (from diagnosis to the end of follow-up)
was 1,346 years and the median follow-up period (DG_to_
    
 
patients (15%) were followed for a period longer than 10
years. The cumulative length of follow-up from the start

and the median follow-up time was 18.5 months.
Study design and data collection
          
DDW on the outcome of breast cancer. We evaluated
232 patients who were continuously enrolled between
       
therapy were administered in 16 hospitals and clinics
in Hungary (listed in the acknowledgements), where

        
Dose (DdU) = 150 (ppm) - D concentration of DDW (ppm) 
body weight (kg)

196
D-concentration and DDW-dose, were recorded. Patient
documentation was retrospectively evaluated and data
collection was closed in January 2012. Patients were
aware of, and provided access to, available information
regarding DDW treatment. Patients were divided into

groups of patients were formed based on the staging at
 74), composed of patients with
advanced breast cancer with distant metastasis or locally
advanced disease, and Group E1 (n158), composed of
patients with early stage breast cancer whose disease did

time point was when patients entered the study and
started DDW treatment. By that time a high percentage
of patients had received conventional therapies, which
resulted in complete remission in some cases but most
patients had progressive disease when they started DDW
treatment. Three subgroups were formed based on status
    129), composed
of advanced breast cancer patients; Group E2 (n55),
composed of patients with early-stage breast cancer,
48), composed of patients in remission.
MST calculations were performed both from the time of

Data of 74 breast cancer patients, who were in advanced
stage at the time of diagnosis and therefore had limited
        
response relationship.
Statistical evaluation
The primary endpoint of the study was survival and the
Kaplan-Meier method was applied to calculate survival

Statistical Software, Version 12.3.0. Because the study
was performed retrospectively and patients started DDW
       
was computed both from the time of diagnosis (MST from
DG) of breast cancer and from the start of DDW treatment
(MST from DDWstart). The dose-response relationship
in the subgroup of breast cancer patients with advanced
disease was tested using one-way analysis of variance
Table 1 Descriptive statistics of 232 breast cancer patients who underwent DDW treatment in addition to conventional therapies
Variable Median Average SD Cumulative time
Age 50 y 51 y 11 y 
Dg_to_DDWstart 13.2 m 35.8 m 53.6 m 692.8 y
LoDDW 14.1 m 24.5 m 29.8 m 474.2 y
 49.4 m 69.7 m 62.3 m 1346 y
 18.5 m 33.8 m 38.4 m 654 y
Abbreviations: 

(ANOVA) and the Bonferroni post hoc test. The mean

Results
Median survival in breast cancer patients undergoing DDW
treatment in addition to conventional treatment regimens
We calculated the MST of all enrolled breast cancer
patients using the Kaplan-Meier method although we
were aware of the heterogeneity of the study population
regarding staging both at the time of diagnosis and at
the start of DDW treatment. The overall survival time
was 148 months (12.3 years) from the diagnosis of
breast cancer and 89 months (7.4 years) from the start
of DDW treatment. To determine whether these MSTs
are noteworthy, a literature search was performed to
determine the stage of breast cancer that results in
MSTs of the same length. We found that the chance of
long-term survival increases primarily in the subset of
patients whose disease is detected at an early stage [17].
        
breast cancer patients enrolled in the present study were
in the most advanced stage of disease at start of DDW
treatment, we presume that DDW treatment contributed

Evaluation of the subgroups showed that according to

early-stage breast cancer achieved an MST of 217 months
(18.1 years), compared with 52 months (4.3 years) for
other patients with advanced cancer. MSTs from the start
     
       
group of patients with early stage breast cancer, while
patients in advanced stage achieved an MST of 39 months
(3.3 years). It is outstanding that MST calculations are
still pending in patients who started DDW treatment in
remission; only one out of these 48 patients died during
    
        
     
underlying the fact that staging is a major determining
factor in the survival of breast cancer. Moreover, all these


197
Table 2 
Response N Mean Std. Deviation

Minimum Maximum
Lower Bound Upper Bound
 16 1.6879 1.02796 1.1401 2.2357 0.45 4.71
 29 1.2897 0.57248 1.0720 1.5075 0.20 2.36
 12 0.6638 0.35056 0.4411 0.8865 0.20 1.17
PD 28 0.9289 0.66529 0.6709 1.1868 0.02 2.27
Total 85 1.1574 0.75377 0.9949 1.3200 0.02 4.71
Abbreviations: 
PD = progressive disease
Figure 1 Kaplan-Meier graphs of survival from the start of DDW

were 39 months for group A2 (n=129) and 89 months for group E2
    
one death in a 221.1-year cumulative follow-up period was recorded, p
(between groups) <0.0001.
Abbreviations:
= patients in remission; DDWstart = start of DDW treatment.
MSTs are long in comparison to other clinical studies on
breast cancer outcome.
Analysis of patients with advanced disease at the start of
DDW treatment
       
   

the subgroup of 74 advanced cancer patients comparing
to historical control [4, 5]. Descriptive statistics of the
subgroup showed that the median time from diagnosis of
advanced stage to the start of DDW treatment was 181
days (5.9 months), with a standard deviation (SD) of 424.5
days (13.9 months). The median length of DDW treatment
(LoDDW) was 402 days (13.2 months), with SD of 569.26
days (18.7 months).
        
to combined treatment with DDW and conventional
therapies

therapies, 16 cases (21.6%) showed a complete response

        
progressive disease (PD) was recorded in 19 (25.7%).
Using the Kaplan-Meier method, MSTs from the start of



between MSTs and the response to DDW therapy.
Figure 2 Kaplan-Meier graphs of survival from the start of DDW

        

months (n=19) p (between groups) <0.0001.
Abbreviations:
change; PD = progressive disease.

Analysis of the relationship between DDW-dose and
response to DDW treatment in breast cancer patients at an
advanced stage
Preclinical studies suggested that the main determining
         
dose of DDW (DdU). The DdU was in the range of 0.02
to 4.71 in the population during the follow-up, and
the response to DDW treatment was determined at 85
time points in the 74 patients. Table 2 shows the means
      

198
The highest m   
   

the standard deviations between the subgroups, one-way
analysis of variance (ANOVA) and the Bonferroni post hoc
  
between the subgroups (p0.001) using the one-way
ANOVA test. Analysis of certain subgroup pairs revealed

 0.004).
The results suggest that higher DDW-dose results in a
pronounced response to DDW treatment.
Analysis of the effect of early- or late-onset of DDW
treatment after diagnosis of breast cancer
In the present retrospective trial the enrollment of
patients was continuous and the time period between
diagnosis and DDW start was relatively long in most
patients (median 13.2 months). Therefore, we investigated
whether the length of time from diagnosis to the start of
       
treatment on the outcome of the disease. Patients were
 114) contained
patients that joined the study within 1 year after their
illness was diagnosed, and group II (n118) contained
patients who started DDW treatment more than 1 year

trial a higher proportion of patients in group II (79.7%)
were staged at advanced disease compared with those in
group I (30.7%). Early stage breast cancer was diagnosed
in 36.8% of the patients in group I and in 11% of group
II. On the other hand, the number of patients who were
successfully treated and achieved remission due to

II than in group I (9.3% versus 31.6%).
The MST for group I (n114) is still pending although
18 deaths were registered during the cumulative follow-
up period of 399.8 years, which indicates 1 death per
22.2 years. The MST for group II was 49 months (4.1
   majority of
      

survival period in this subset of patients.
        
treatments: Single versus repeated DDW treatment
      
metastases is a crucial aspect of breast cancer, even

      
frequency of DDW treatment. A single DDW treatment
was administered to 179 patients (group N) whereas 53
patients repeated the cure at least once (group Y). The
Kaplan-Meier graphs show that the survival curves have

DDW at least twice had an MST of 293 months (24.4 years)
whereas those who received a single DDW treatment had
an MST of 108 months (9 years).
Figure 3 Kaplan-Meier graphs of survival from start of DDW treatment
           

patients), cumulative follow-up was 399.8 years, indicating one death
in 22.2 years. Group II (118 patients) showed a MST of 49 months, p
(between groups) <0.0001.
Abbreviation: DG_to_DDWstart = time from the diagnosis to the start of
DDW treatment.
Figure 4 Kaplan-Meier graphs of survival from the diagnosis in breast
cancer patients undergoing single (group N) or repeated (group Y) DDW
 
months, compared with 108 months for group N (n=179 patients, single
DDW treatment), p (between groups) <0.0001.
Survival in breast cancer patients enrolled before and after
January 2005
The present retrospective study covers a wide interval
from April 1993 to January 2012. During that time there
       
cancer. The entire study population was divided into two
subgroups based on the date of involvement so that we
could investigate, whether D-depletion contributed to
the longer survival, when it was applied in addition to
the recent treatment regimens. The subgroup of patients
who entered the study before January 2005 (n156)
showed longer survival in comparison to the historical
controls. The MST was 71 months (5.9 years) in the group
of patients with early stage (n34) breast cancer, while


199
patients in advanced stage (n86) achieved an MST of
        
still pending in patients who started DDW treatment in
remission; only one out of these 36 patients died. MSTs
in the subgroup of patients enrolled after January 2005
was non-calculable. Survival from start of DDW treatment

cumulative follow-up period of 118.7 years, despite of the
fact, that high proportion of these patients was diagnosed
with advanced breast cancer (n
from primary disease and only 12 patients were in
remission.
Figure 5 Kaplan Meier graphs of survival from the start of DDW

for group A2 (n=86) and 71 months for group E2 (n=34). The MST for
  
(between groups) <0.0001.
Abbreviations:
= patients in remission; DDWstart = start of DDW treatment.
Discussion
  and in vivo   
that naturally occurring deuterium plays a pivotal role
in cell growth and a shortage of deuterium in tumour
cells induces apoptosis, resulting in partial or complete

      
of D-depletion into conventional treatments delayed

prevention of relapses [14, 19], and reduced the risk of
  
[13]. To gain further information on the application of
D-depletion, we retrospectively evaluated records of 232
breast cancer patients who underwent DDW treatment in
addition to conventional treatment regimens.
The overall survival rates calculated in the present study
suggest that DDW treatment noticeably prolonged MST
in comparison with published data [5, 17]. The MST of
the entire cohort was 148 months from diagnosis of the
disease and 89 months from the start of DDW treatment.

showed that MST correlated with the staging of breast
cancer [4, 17] with an MST of 217 months in patients with
early disease and 52 months in advanced cases. We were
aware that the study population was heterogeneous. All
medical records were thoroughly investigated for possible
       
subgroup evaluation was performed based on these
aspects. It is well known that retrospective approach has
limitations but one of the advantages of a retrospective
study is their usefulness in addressing rare conditions and
it helps to assess the feasibility of prospective studies.
In Group I patients (n114), who started DDW treatment
within 1 year after diagnosis, only 18 deaths were
registered (one death in every 22.2 years). The low death
rate did not allow the calculation of MST. When DDW
treatment was started later than 1 year after diagnosis
(Group II, n118) the MST was 49 months despite the
fact that 79.7% of the patients were in advanced stage.
The cumulative follow-up of 1,346 years in the present
study allowed us to evaluate whether D-depletion was
     
      
         

was also applicable to patients who were in remission
after successful conventional treatment (n48), and the
cumulative follow-up period and reported deaths indicated
one death per 221.1 years, which is an outstanding result
  

subpopulation.
Evaluation of the correlation between DDW-dose (DdU)
and response to treatment provided evidence on the
       
patients whose disease did not change or progressed
         
Additionally, irregular consumption resulted in alterations
        


These facts strongly suggest that in most cases the failure
of DDW therapy could be attributed to improper dosage


    
(52 months) were two or three times longer than those of
patients who received only conventional treatment (MST,
12-31 months).
       
D-depletion was the frequency of DDW applications
(single or repeated treatment). Although single DDW
treatment (n    
with published MSTs, repeated DDW treatment resulted
in outstandingly long survival periods. Patients who took
DDW for only one treatment period (n126) had an MST
of 9 years whereas patients (n=53) who took DDW more
than once attained an MST of 24.4 years.


200
It is crucial to investigate survival data of breast cancer
patients from the point of long-term follow-up, because
long-term survival and the chance for late onset of
recurrence are also characteristic for breast cancer. There
are advances in the knowledge about breast cancer, in its
molecular basis, in early detection and multidisciplinary
treatment options. Our study covers a wide time interval,
and this fact raises the question, whether D-depletion
         
patients undergoing the most recent treatment regimens.
        
noticeably longer compared to historical control, and did

       


support the evidence that DDW treatment contributed
        
        

Our data indicate that repeated DDW treatment is highly

to longer survival and possibly the prevention of disease
      
        
conventional therapies noticeably prolonged MST in
breast cancer patients. The above results are impressive
in breast cancer research. All this information is useful
in conduction of further prospective trials to clarify the

Conclusion

treatment of early-stage or advanced breast cancer and
in preventing disease recurrence. The method is safe and

       
absolutely no adverse events occurred during long-term
application of DDW at a wide concentration range of 25 to

       
into standard treatment regimens for breast cancer.
Acknowledgements
The authors wish to acknowledge the clinicians who
carried out the conventional forms of cancer therapy
     
   
     
Budapest; Korányi National Institute of Tuberculosis
and Pulmonology, Budapest; National Institute of
Oncology, Budapest; Péterfy Sándor Hospital, Budapest;
     
Hospital, Budapest; St. István Hospital, Budapest; St.
     
     


Hospital, Berettyóújfalu.


interest.
References
 

         
   

.
        


       
Overview of resistance to systemic therapy in patients with breast

   

          


          

       
       

[9] Somlyai G, Jancsó G, Jákli G, Vass K, Barna B, et al. (1993) Naturally
occurring deuterium is essential for the normal growth rate of cells.

[10] Somlyai G, Laskay G, Berkényi T, JákliGy, Jancsó G (1998) Naturally
occurring deuterium may have a central role in cell signalling.
         
     

[11] Somlyai G, Laskay G, Berkényi T, Galbács Z, Galbács G, et al. (1998)


[12] Gyöngyi Z, Somlyai G (2000) Deuterium depletion can decrease the
        




[14] Kovács A, Guller I, Krempels K, Somlyai I, Jánosi I, Z, et al. (2011)
Deuterium depletion may delay the progression of prostate cancer.


         


      
          


(2008) An overview of prognostic factors for long-term survivors of

            
Deuterium-depleted water inhibits human lung carcinoma cell

[19] Krempels K, Somlyai I, Somlyai G (2008) A retrospective evaluation
         
        

           
        

